oue ssue Assayise Letters - AAT Bioquest · 2/1/2013 · atiometrR uri F a Caium ndiaI orst 1 onadioatie uose ptake Assays ... all products are for Research Use Only. ... 21050 Quin-2

New Product HighlightsRatiometric Fura-6™ Calcium IndicatorsNon-Radioactive Glucose Uptake AssaysMulticolor Cell Viability Assays for Flow Cytometry

Biochemical AssaysColorimetric Quantitation of AmmoniaFluorimetric Quantitation of Formaldehyde Fluorimetric Quantitation of Lactate DehydrogenaseFluorimetric Measurement of Glucose-6-Phosphate Detection of Reactive Oxygen Species (ROS)

Cell-Based AssaysFluo-8® Green Calcium IndicatorsGaussia Luciferase Reporter Gene Assay Firefly Luciferase Reporter Gene AssayiFluor™ 594, a Bright Fluorescent Labeling Dye

Labeling ProbestrFluor™ Eu TRF Labeling DyestrFluor™ Tb TRF Labeling DyesReadiLinkTM iFluorTM Antibody Labeling Kits

2013 • Volume 2 • Issue 1

AssayWise Letters

AAT Bioquest®

Advancing Assay & Test Technologies

Table of Contents

New Product Highlights

Ratiometric Fura-6™ Calcium Indicators 1

Non-Radioactive Glucose Uptake Assays 2

Multicolor Cell Viability Assays for Flow Cytometry 3

Biochemical Assays

Colorimetric Quantitation of Ammonia 4

Fluorimetric Quantitation of Formaldehyde 4

Fluorimetric Quantitation of Lactate Dehydrogenase 5

Fluorimetric Measurement of Glucose-6-Phosphate 5

Detection of Reactive Oxygen Species (ROS) 6

Cell-Based Assays Fluo-8® Green Calcium Indicators 7

Gaussia Luciferase Reporter Gene Assay 9

Firefly Luciferase Reporter Gene Assay 9

iFluor™ 594, a Bright Fluorescent Labeling Dye 10

Labeling Probes

trFluor™ Eu TRF Labeling Dye 12

trFluor™ Tb TRF Labeling Dye 12

ReadiLink™ iFluor™ Antibody Labeling Kits 13 Trademarks of Other Companies

Alexa Fluor® (Invitrogen)Cy5® (GE Healthcare)DyLight™ (ThermoFisher)

FlexStation® (Molecular Devices)SpectrumMax® (Molecular Devices)

Trademarks of AAT Bioquest

AAT Bioquest®Amplite™ Cal-520™Cell Explorer™Fluo-8®Fluo-8FF™Fluo-8H™Fluo-8L™Fura-6™

iFluor™ mFluor™ReadiLink™ReadiView™ROS Brite™Screen Quest™Tide Quencher™trFluor™

AAT Bioquest, Inc. (formerly ABD Bioquest, Inc.) develops, manufactures and markets bioanalytical research reagents and kits to life sciences, diagnostic R&D and drug discovery. We specialize in photometric detections including absorption (color), fluorescence and luminescence technologies. AAT Bioquest offers a rapidly expanding list of enabling products. AssayWise Letters is a platform for AAT Bioquest to introduce its newest products and services, and to update the new applications of our existing products. The Company's superior products enable life science researchers to better understand biochemistry, immunology, cell biology and molecular biology. AAT Bioquest also offers custom service to meet the distinct needs of each customer.

It is my greatest pleasure to welcome you to this new issue of our AssayWise Letters. While we continue to rapidly expand, our core value remains the same: Innovation and Customer Satisfaction. We are commit-ted to being the provider of novel biological detection solutions. We promise you to extend these values to you during the course of our service and to continue to support you with our new products and services. It is our greatest honor to receive valuable feedback and sugges-tions from you.

Very truly yours,

Zhenjun Diwu, Ph.D.President

From the President of AAT Bioquest

www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304 New Product Highlights

2013, 2 (1) • AssayWise LettersUnless otherwise specified, all products are for Research Use Only. Not for use in diagnostic or therapeutic procedures. 1

Ratiometric Fura-6™ Calcium Indicators

Although Fura-2 has been widely used as the preferred excitation-ratioable calcium indicator, it has certain limitations, e.g., lower sensitivity compared to the single wavelength calcium dyes such as Fluo-8® and Cal-520™. AAT Bioquest has recently developed Fura-6™ to improve the calcium response of Fura-2. As demonstrated in Figures 1.1 and 1.2, Fura-6™ AM is more sensitive to calcium than Fura-2 AM. In addition, Fura-6™ has its emission shifted to longer wavelength (Em = 525 nm). Fura-6™ might be also used for the flow cytometric analysis of calcium in cells due to its excellent excitation at 405 nm that perfectly matches the violet laser line of flow cytometer.

Key Features of Fura-6™

• Fura-6™ responses to calcium the same way as Fura-2 does

• Red-shifted dual excitation wavelengths (354 nm/415 nm)

• Better excited at 405 nm for flow cytometric applications

• Compatible with common filter sets

• Higher signal/background ratio than that of Fura-2

Figure 1.1. Fluorescence excitation spectra of Fura-6™ in the presence of 0 to 39 µM free Ca2+.

Figure 1.2. ATP dose responses in CHO-K1 cells measured with Fura-2 AM and Fura-6™ AM respectively. CHO-K1 cells were seeded overnight at 40,000 cells/100 μL/well in a Costar black wall/clear bottom 96-well plate. The cells were incubated with Fura-2 AM and Fura-6™ AM calcium assay dye-loading solution respectively for 1 hour at room temperature. ATP (50 μL/well) was added by FlexStation®.

Fura-2 AM

Fura-6™ AM

Table 1.1. Product Ordering Information

Cat. # Product Description SizeZero Calcium High Calcium

Kd (nM)Ex (nm) Em (nm) Ex (nm) Em (nm)

21054 BTC AM 1 mg 464 533 401 529 7,000

21053 BTC, tetrapotassium salt 1 mg 464 533 401 529 7,000

21021 Fura-2 AM *UltraPure grade* 1 mg 363 512 335 505 145

21025 Fura-2, pentapotassium salt 1 mg 363 512 335 505 145

21026 Fura-2, pentasodium salt 1 mg 363 512 335 505 145

21055 Fura-6™ AM 1 mg 386 532 354 524 260

21056 Fura-6™ AM 10x50 μg 386 532 354 524 260

21057 Fura-6™, potassium salt 1 mg 386 532 354 524 260

21058 Fura-6™, sodium salt 1 mg 386 532 354 524 260

21032 Indo-1 AM *UltraPure grade* 1 mg 346 475 330 401 230

21040 Indo-1, pentapotassium salt 1 mg 346 475 330 401 230

21044 Indo-1, pentasodium salt 1 mg 346 475 330 401 230

21050 Quin-2 AM 1 mg 353 495 333 495 60

21052 Quin-2, tetrapotassium salt 5 mg 353 495 333 495 60


Cat. # Product Description Size Ex (nm) Em (nm)22600 Cell Explorer™ Fixable Cell Viability Assay Kit *Blue Fluorescence* 1 kit 353 442

22500 Cell Explorer™ Fixable Cell Viability Assay Kit *Blue Fluorescence with 405 nm Excitation* 1 kit 410 450

22604 Cell Explorer™ Fixable Cell Viability Assay Kit *Deep Red Fluorescence* 1 kit 649 660

22601 Cell Explorer™ Fixable Cell Viability Assay Kit *Green Fluorescence* 1 kit 498 521

22501 Cell Explorer™ Fixable Cell Viability Assay Kit *Green Fluorescence with 405 nm Excitation* 1 kit 408 512

22602 Cell Explorer™ Fixable Cell Viability Assay Kit *Orange Fluorescence* 1 kit 547 573

22502 Cell Explorer™ Fixable Cell Viability Assay Kit *Orange Fluorescence with 405 nm Excitation* 1 kit 398 550

22603 Cell Explorer™ Fixable Cell Viability Assay Kit *Red Fluorescence* 1 kit 583 603

Cat. # Product Description Size Ex (nm) Em (nm)11300 Amplite™ Fluorimetric Glucose Oxidase Assay Kit *Red Fluorescence* 1 kit 571 585

40005 Amplite™ Glucose Quantitation Kit 1 kit 571 585

11705 Glucose-UDP-Fluorescein Conjugate 100 μg 490 514

11706 Glucose-UDP-(PEG)6-Fluorescein Conjugate 100 μg 490 514

36503 Screen Quest™ Colorimetric Glucose Uptake Assay Kit 1 kit 575 N/A

36500 Screen Quest™ Fluorimetric Glucose Uptake Assay Kit 1 kit 571 585


2 Unless otherwise specified, all products are for Research Use Only. Not for use in diagnostic or therapeutic procedures.

New Product Highlights www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304

Multicolor Cell Viability Assays for Flow Cytometry

Cell Explorer™ fluorescence labeling kits are a set of tools to label cells for fluorescence microscopic and flow cytometric investiga-tions of cellular functions. AAT Bioquest offers a set of multicolor cell labeling kits to uniformly label dead mammalian cells for long term microscopic examination and flow cytometric analysis of cells.

The kits provide all the essential components with an optimized cell-labeling protocol. The proprietary fluorescent dyes used in the kits become more fluorescent upon reacting with dead cellular components. When excited at different excitations using a set of our Cell Explorer™ cell imaging kits, the dead cell stain emits in-tense fluorescence ranging from 400 nm to 800 nm. The fluorescent dyes used in the kits have the spectral properties compatible with all the common filter sets. The kits can be readily used with almost every fluorescence instrument often equipped with 405 nm violet laser, 488 nm blue laser or 633 nm red laser sources.

Figure 1.6. Jurkat cells were treated and stained with Cell Explorer™ Fixable Cell Viability Assay Kits 22601 (Panel A) and 22604 (Panel B). The cells were fixed in 3.7% formaldehyde and analyzed by flow cytometry. Live (blue solid peak), staurosporine treated (green line) and heat-treated (red solid peak) cells were distinguished with Ex/Em = 488 nm/520 nm (FL1) channel (panel A) and 633 nm/660 nm (FL4) channel (Panel B). Nearly identical results were obtained using unfixed cells.

Unless otherwise specified, all products are for Research Use Only. Not for use in diagnostic or therapeutic procedures. 3

www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304 New Product Highlights

2013, 2 (1) • AssayWise LettersAssayWise Letters • 2013, 2 (1)

Figure 1.4. 2-DG uptake in differentialed 3T3-L1 adipocytes and 3T3-L1 fibroblasts. Assays were performed with Screen Quest™ Fluorimetric Glucose Uptake Assay Kit (Cat# 36500) in a black wall/clear bottom cell culture Poly-D-Lysine plate using a Gemini (Molecular Devices) microplate reader. A: Negative Control, no insulin and no 2-DG treatment; B: 2-DG uptake in the absence of insulin; C: 2-DG uptake in the presence of 1 μM insulin; D: 2-DG uptake in the presence of 1 μM insulin and 200 μM phloretin; E: 2-DG uptake in the presence of 1 μM insulin and 5 mM D-glucose.

Non-Radioactive Glucose Uptake Assays

Figure 1.5. Image of CHO cells fixed with formaldehyde and stained with Cell Explorer™ Fixable Cell Viability Assay Kit (Cat# 22601) in a Costar 96-well black wall/clear bottom plate.

Glucose transport systems are responsible for transporting glucose across cell membranes. Measuring the uptake of 2-deoxyglucose (2-DG), a glucose analog, is widely accepted as a reliable method to estimate the amount of glucose uptake and to investigate the regulation of glucose metabolism and insulin resistance. The 2-DG uptake is commonly determined using the non-metabolized 2-DG labeled with tritium or C14. However, the use of a radiolabeled probe is costly and requires a tedious special handling procedure.

AAT Bioquest’s Screen Quest™ Colorimetric Glucose Uptake Assay Kit provides a sensitive and non-radioactive glucose uptake assay. In this assay 2-DG is taken up by glucose transporters, and metabo-lized to 2-DG-6-phosphate (2-DG6P). The non-metabolizable

Figure 1.3. Glucose transportation into cells via glucose transporters, Glu 1, Glu 2, Glu 3 and Glu 4.

A

B

2-DG6P accumulates in cells and is proportional to glucose uptake. The accumulated 2-DG6P is enzymatically coupled to generate NADPH, which is specifically monitored by a chromogenic NADPH sensor. The signal can be read using an absorption microplate reader by reading the OD ratio of 570 nm to 610 nm.

Based on the same principle, AAT Bioquest’s Screen Quest™ Fluorimetric Glucose Uptake Assay Kit provides an even more sensitive and non-radioactive glucose uptake assay in tissues or cultured cells. The accumulated 2-DG6P is enzymatically coupled to generate NADPH, which is specifically monitored by a fluorogenic NADPH sensor. The signal can be read using a fluorescence microplate reader at Ex/Em = 540 nm/590 nm.


Cat. # Product Description Size Ex (nm) Em (nm)10051 Amplite™ Colorimetric Aldehyde Quantitation Kit 1 kit 550 N/A

10053 Amplite™ Colorimetric Aldehyde Quantitation Kit *Blue Color* 1 kit 620 N/A

10059 Amplite™ Colorimetric Ammonia Quantitation Kit *Blue Color* 1 kit 650 N/A

10058 Amplite™ Colorimetric Urea Quantitation Kit *Blue Color* 1 kit 650 N/A

10052 Amplite™ Fluorimetric Aldehyde Quantitation Kit 1 kit 360 450

10057 Amplite™ Fluorimetric Formaldehyde Quantitation Kit *Green Fluorescence* 1 kit 400 510

Ammonia is an important source of nitrogen for living systems. It is produced in liver and converted to urea through the urea cycle. Ammonia is synthesized through amino acid metabolism and is toxic when present at high concentrations. Elevated levels of ammonia in blood (hyperammonemia) have been found in liver dysfunction (cirrhosis), while hyperammonemia is associated with defects in the urea cycle enzymes (e.g. ornithine transcarbamylase). The determination of ammonia is very useful for clinical laboratory to monitor health status.

AAT Bioquest’s Amplite™ Colorimetric Ammonia Quantitation Kit provides a simple and sensitive colorimetric method for the quantitation of ammonia in foods and biological samples such as serum, plasma and urine, etc. The assay is based on an enzyme-coupled reaction of ammonia, which generates a blue colored product. The absorbance of the blue product is proportional to the concentration of ammonia, which can be measured colorimetrically at 660-670 nm. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format.

Colorimetric Quantitation of Ammonia

Figure 2.1. Ammonia dose responses were measured in a 96-well clear bottom plate with Amplite™ Colorimetric Ammonia Quantitation Kit. As low as 4 μM ammonia can be detected (n=3) with 45 minute incubation.

Figure 2.2. Formaldehyde dose responses were measured in a 96-well black plate with Amplite™ Fluorimetric Formalhyde Quantitation Kit. As low as 1 µM formaldehyde can be detected with 30 minute incubation (n=3).

Formaldehyde is a naturally occurring substance. Natural processes in the upper atmosphere may contribute up to 90 percent of the total formaldehyde in the environment. Formaldehyde, as well as its oligomers and hydrates are rarely encountered in living organisms. Methanogenesis proceeds via the equivalent of formaldehyde, but this one-carbon species is masked as a methylene group in methanopterin. Formaldehyde is the primary cause of methanol’s toxicity, since methanol is metabolized into toxic formaldehyde by alcohol dehydrogenase.

AAT Bioquest's Amplite™ Fluorimetric Formaldehyde Quantitation Kit is used for quantifying formaldehyde. The kit uses a proprietary fluorogenic dye that generates a strongly fluorescent product upon reacting with formaldehyde. This fluorimetric kit provides a sensi-tive mix-and-read method to detect formaldehyde. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. Its signal can be easily read using a fluorescence microplate reader at Ex/Em = 400/510 nm.

Fluorimetric Quantitation of Formaldehyde

Lactate dehydrogenase (LDH) is an oxidoreductase enzyme that catalyzes the interconversion of pyruvate and lactate. Localized in the cytosol, LDH is present in a wide variety of organisms, including animals and plants. Cells release LDH into the bloodstream after tissue damage or red blood cell hemolysis. Since LDH is a fairly stable enzyme, it has been widely used to evaluate the presence of damage and toxicity of tissue and cells. Quantification of LDH has a broad range of applications.

AAT Bioquest’s Amplite™ Lactate Dehydrogenase Assay Kits provide both fluorescence- and absorbance-based methods for detecting either L-lactate dehydrogenase (L-LDH) or D-lactate dehydrogenase (D-LDH). In the enzyme coupled assay, LDH is proportionally related to the concentration of NADH that is specifically monitored using a fluorogenic NADH sensor. The fluorescence signal can be read using a fluorescence microplate reader at Ex/Em = 540 nm/590 nm. We were able to detect as low as 1 mU/mL L-LDH in a 100 μL reaction volume. The assays are robust, and can be readily adapted for a wide variety of applications that require the measure-ment of L-LDH.

Fluorimetric Quantitation of Lactate Dehydrogenase

Figure 2.3. L-LDH dose responses were measured with Amplite™ Fluorimetric L-LDH Assay Kit in a 96-well black plate. As low as 1 mU/mL L-LDH in a 100 μL reaction volume can be detected with 30 minute incubation.

Glucose-6-phosphate (G6P) is a key intermediate for glucose to transport into cells. G6P may also be converted to glycogen or starch for storage in the liver and muscles. G6P is utilized by glucose-6-phosphate dehydrogenase (G6PD) to generate the reducing equivalents in the form of NADPH. This is particularly important in red blood cells where G6PD deficiency leads to hemolytic anemia.

AAT Bioquest’s Amplite™ Fluorimetric Glucose-6-Phosphate Assay Kit provides a simple, sensitive and rapid fluorescence-based method for detecting G6P in biological samples such as serum, plasma, urine, as well as in cell culture samples. In the coupled enzyme assay, the G6P concentration is proportionally related to NADPH that is specifically monitored by a fluorogenic NADPH sensor. The fluorescence signal can be read by a fluorescence microplate reader at Ex/Em = 530-570 nm/590-600 nm (Ex/Em = 540 nm/590 nm is recommended). With the Amplite™ G6P Assay Kit, we were able to detect as low as 0.3 μM G6P in a 100 μL reaction volume.

Fluorimetric Measurement of Glucose-6-Phosphate

Figure 2.4. G6P dose responses were measured with Amplite™ Fluorimetric G6P Assay Kit in a 96-well black plate using a Gemini (Molecular Devices) microplate reader. As low as 0.3 μM G6P in a 100 μL reaction volume can be detected with 1 hour incubation.

4 Unless otherwise specified, all products are for Research Use Only. Not for use in diagnostic or therapeutic procedures.AssayWise Letters • 2013, 2 (1)

Biochemical Assays www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304

www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304 Biochemical Assays

Unless otherwise specified, all products are for Research Use Only. Not for use in diagnostic or therapeutic procedures. 5 2013, 2 (1) • AssayWise Letters


Cat. # Product Description Size Ex (nm) Em (nm)13809 Amplite™ Colorimetric D-Lactate Dehydrogenase (LDH) Assay Kit 1 kit 575 N/A

13807 Amplite™ Colorimetric Glucose-6-Phosphate Dehydrogenase (G6PD) Assay Kit 1 kit 575 N/A

13813 Amplite™ Colorimetric L-Lactate Dehydrogenase (LDH) Assay Kit 1 kit 575 N/A

13808 Amplite™ Fluorimetric D-Lactate Dehydrogenase (LDH) Assay Kit 1 kit 571 585

13804 Amplite™ Fluorimetric Glucose-6-Phosphate Assay Kit 1 kit 571 585

13806 Amplite™ Fluorimetric Glucose-6-Phosphate Dehydrogenase (G6PD) Assay Kit 1 kit 571 585

13812 Amplite™ Fluorimetric L-Lactate Dehydrogenase (LDH) Assay Kit 1 kit 571 585

Reactive oxygen species (ROS) are chemically reactive molecules containing oxygen (such as superoxide, hydroxyl radical, singlet oxygen and peroxides). ROS is highly reactive due to the presence of unpaired valence shell electrons. ROS forms as a natural byproduct of the normal metabolism of oxygen and plays important roles in cell signaling and homeostasis. However, during times of environmental stress (e.g., UV or heat exposure), ROS levels can increase dramatically. It may result in significant damage to cell structures. Cumulatively, this is known as oxidative stress. ROS is also generated by exogenous sources such as ionizing radia-tion. Under the conditions of oxidative stress, greatly increased production of ROS results in subsequent alteration of membrane lipids, proteins and nucleic acids. Oxidative damage of these biomolecules is associated with aging as well as with a variety of pathological events, including atherosclerosis, carcinogenesis, ischemic reperfusion injury, and neurodegenerative disorders.

ROS Brite™ reagents are a series of new fluorogenic probes to measure oxidative stress in cells. The cell-permeant ROS Brite™ reagents are nonfluorescent and produce bright fluorescence upon ROS oxidation. The resulting fluorescence can be measured using fluorescence imaging, high-content imaging, microplate fluorometry, or flow cytometry. ROS Brite™ 570, 670 and 700 reagents have good selectivity to both hydroxyl radical and supoeroxide.

ROS Brite™ 570 reagent is a new fluorogenic probe to measure oxi-dative stress in cells using conventional fluorescence microscopy, high-content imaging, microplate fluorometry, or flow cytometry. The cell-permeant ROS Brite™ 570 reagent is nonfluorescent and produces bright orange fluorescence upon ROS oxidation.

ROS Brite™ 670 can be well excited with He-Ne laser at 633 nm, making this reagent well suited for the ROS detection using a flow cytometer. Its fluorescence signal can be well monitored using the Cy5® filter set.

ROS Brite™ 700 reagent is a new fluorogenic probe to measure oxidative stress in small animals. The cell-impermeant ROS Brite™ 700 reagent is water-soluble nonfluorescent and produces bright near-infrared fluorescence upon ROS oxidation. The resulting fluorescence can be measured using in vivo fluorescence imaging.

Detection of Reactive Oxygen Species (ROS)

Using Multicolor ROS BriteTM Reagents

Figure 2.5. The fluorescence spectra of ROS Brite™ 570, ROS Brite™ 670 and ROS Brite™ 700 in PBS buffer (pH 7.2).

Figure 2.6. The responses of ROS Brite™ 570 to different ROS species.

Fluo-8® Green Calcium IndicatorsThe Brightest Ca2+ Signal at 488 nm Excitation

Fluo-3 and Fluo-4 were the most commonly used visible light-excitable calcium indicators. However, Fluo-3 AM and Fluo-4 AM are only moderately fluorescent in live cells upon esterase hydrolysis, and require harsh cell loading conditions to maximize their cellular calcium responses. Fluo-8® dyes have been developed to improve cell loading and calcium response while maintaining the convenient Fluo-3 and Fluo-4 spectral wavelengths of maximum excitation @ ~490 nm and maximum emission @ ~520 nm. For cell loading, Fluo-8® AM only requires incubation at room temperature while Fluo-3 AM and Fluo-4 AM require incubation at 37 oC. In addition, Fluo-8® AM is 2 times brighter than Fluo-4 AM, and 4 times brighter than Fluo-3 AM in cells. AAT Bioquest offers a set of outstanding Fluo-8® reagents with different calcium binding affinities.

Key Features of Fluo-8® AM

• Faster, more readily loaded into cells than Fluo-3 AM and

Fluo-4 AM. Only room temperature is required.

• Brighter, much brighter than Fluo-3 AM and Fluo-4 AM in cells.

• Convenient, almost identical spectra to those of Fluo-4 AM.

Figure 3.1. Carbachol dose responses were measured in HEK-293 cells with Fluo-8® AM and Fluo-4 AM. HEK-293 cells were seeded overnight at 40,000 cells/100 µL/well in a 96-well black wall/clear bottom Costar plate. The growth medium was removed, and the cells were incubated with 100 µL of dye-loading solution containing Fluo-8® AM or Fluo-4 AM for 1 hour at room temperature. Carbachol (25µL/well) was added by NOVOstar to achieve the final indicated concentrations. The fluorescence signals were measured at Ex/Em = 490/525 nm. The EC50 of Fluo-8® AM is about 1.2 µM.

Carbachol Dose (μM)

Fluo-8® AM

Fluo-4 AM

Figure 3.2. U2OS cells were seeded overnight at 40,000 cells per 100 µL per well in a Costar black wall/clear bottom 96-well plate. The growth medium was removed, and the cells were incubated with 100 µL of 4 µM Fluo-3 AM, Fluo-4 AM and Fluo-8® AM in HHBS at 37 °C for 1 hour. The cells were washed twice with 200 µL HHBS, then imaged with a fluores-cence microscope using FITC channel.

Fluo-3 AM Fluo-4 AM Fluo-8® AM

Cat # Product Description Size Ex (nm) Em (nm) Kd (nM)21080 Fluo-8® AM 1 mg 494 517 389

21081 Fluo-8® AM 5x50 µg 494 517 389

21088 Fluo-8®, sodium salt 10x50 µg 494 517 389

21104 Fluo-8FF™ AM 10x50 µg 494 517 10,000

21102 Fluo-8FF™, potassium salt 10x50 µg 494 517 10,000

21090 Fluo-8H™ AM 1 mg 494 517 232

21095 Fluo-8H™, sodium salt 10x50 µg 494 517 232

21096 Fluo-8L™ AM 1 mg 494 517 1,860

21098 Fluo-8L™, sodium salt 10x50 µg 494 517 1,860


Biochemical Assays www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304

6 Unless otherwise specified, all products are for Research Use Only. Not for use in diagnostic or therapeutic procedures.AssayWise Letters • 2013, 2 (1) Unless otherwise specified, all products are for Research Use Only.

Not for use in diagnostic or therapeutic procedures. 7 2013, 2 (1) • AssayWise Letters

www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304 Cell-Based Assays


Cat. # Product Description Size Ex (nm) Em (nm)

16000 ROS Brite™ 570 1 mg 556 566

16002 ROS Brite™ 670 1 mg 650 666

16004 ROS Brite™ 700 *Optimized for in vivo Imaging* 1 mg 680 706

16050 ROS Brite™ APF 1 mg 492 515

16051 ROS Brite™ HPF 1 mg 492 515

Recent Citations of Fluo-8® Calcium Indicators

André M. Deslauriers, Amir Afkhami-Goli, Amber M. Paul, Rakesh K. Bhat, Shaona Acharjee, Kristofor K. Ellestad, Farshid Noorbakhsh, Marek Michalak, and Christopher Power. Neuroinflammation and Endoplasmic Reticulum Stress Are Coregulated by Crocin To Prevent Demyelination and Neurodegeneration. J. Immunol. 2011; 187: 4788 -4799.

Christopher A. Del Negro, John A. Hayes, and Jens C. Rekling. Den-dritic Calcium Activity Precedes Inspiratory Bursts in preBötzinger Complex Neurons. J. Neurosci. 2011; 31: 1017 - 1022.

Ferdinand Maingat, Brendan Halloran, Shaona Acharjee, Guido van Marle, Deirdre Church, M. John Gill, Richard R. E. Uwiera, Eric A. Cohen, Jon Meddings, Karen Madsen, and Christopher Power. Inflammation and epithelial cell injury in AIDS enteropathy: involvement of endoplasmic reticulum stress. FASEB J. 2011; 25: 2211 - 2220.

Jens C. Rekling, Kristian H. R. Jensen, and Henrik Jahnsen. Spontaneous cluster activity in the inferior olivary nucleus in brainstem slices from postnatal mice. J. Physiol. 2012; 590: 1547 - 1562.

Karin Persson and Jens C. Rekling. Population calcium imaging of spontaneous respiratory and novel motor activity in the facial nucleus and ventral brainstem in newborn mice. J. Physiol. 2011; 589: 2543 - 2558.

Mako Kurogi, Megumi Miyashita, Yuri Emoto, Yoshihiro Kubo, and Osamu Saitoh. Green Tea Polyphenol Epigallocatechin Gallate Acti-vates TRPA1 in an Intestinal Enteroendocrine Cell Line, STC-1. Chem Senses 2012; 37: 167 - 177.

Man Hagiyama, Tadahide Furuno, Yoichiroh Hosokawa, Takanori Iino, Takeshi Ito, Takao Inoue, Mamoru Nakanishi, Yoshinori Mu-rakami, and Akihiko Ito. Enhanced Nerve–Mast Cell Interaction by a Neuronal Short Isoform of Cell Adhesion Molecule-1. J. Immunol. 2011; 186: 5983 - 5992.

Marc Aurel Busche, Xiaowei Chen, Horst A. Henning, Julia Reich-wald, Matthias Staufenbiel, Bert Sakmann, and Arthur Konnerth. Critical role of soluble amyloid-β for early hippocampal hyperactiv-ity in a mouse model of Alzheimer’s disease. PNAS 2012; 109: 8740 - 8745.

Md. Shahidul Islam, Calcium Signaling, in Advances in Experimental Medicine and Biology, Vol. 740, Springer, New York, 2012, pp 44-83.

Michael J. Corey, Coupled Bioluminescence Assays, Methods, Evalu-ations and Applications. Wiley, New York. 2009, PP160-190.

Nicholas B. Last, Elizabeth Rhoades, and Andrew D. Miranker. Islet amyloid polypeptide demonstrates a persistent capacity to disrupt membrane integrity. PNAS 2011; 108: 9460 - 9465.

Nicholas M. Mellen and Deepak Mishra. Functional Anatomical Evidence for Respiratory Rhythmogenic Function of Endogenous Bursters in Rat Medulla. J. Neurosci. 2010; 30: 8383 - 8392.

Satoru Torii, Kentaro Kobayashi, Masayuki Takahashi, Kasumi Katahira, Kenji Goryo, Natsuki Matsushita, Ken-ichi Yasumoto, Yoshiaki Fujii-Kuriyama, and Kazuhiro Sogawa. Magnesium Deficiency Causes Loss of Response to Intermittent Hypoxia in Paraganglion Cells. J. Biol. Chem. 2009; 284: 19077 - 19089.

Shaona Acharjee, Yu Zhu, Ferdinand Maingat, Carlos Pardo, Klaus Ballanyi, Morley D. Hollenberg, and Christopher Power. Proteinase-activated receptor-1 mediates dorsal root ganglion neuronal degeneration in HIV/AIDS. Brain 2011; 134: 3209 - 3221.

Soichi Watanabe, Andre P. Seale, E. Gordon Grau, and Toyoji Kaneko. Stretch-activated cation channel TRPV4 mediates hyposmotically induced prolactin release from prolactin cells of mozambique tila-piaOreochromis mossambicus. Am J Physiol Regulatory Integrative Comp Physiol. 2012; 302: R1004 - R1011.

Takako Saito, Kogiku Shiba, Kazuo Inaba, Lixy Yamada, and Hitoshi Sawada. Self-incompatibility response induced by calcium increase in sperm of the ascidian Ciona intestinalis. PNAS 2012; 109: 4158 - 4162.

Wei-Wei Shen, Maud Frieden, and Nicolas Demaurex. Local Cyto-solic Ca2+Elevations Are Required for Stromal Interaction Molecule 1 (STIM1) De-oligomerization and Termination of Store-operated Ca2+ Entry. J. Biol. Chem.2011; 286: 36448 - 36459.

Wen Fu, Araya Ruangkittisakul, David MacTavish, Jenny Y. Shi, Klaus Ballanyi, and Jack H. Jhamandas. Amyloid β (Aβ) Peptide Directly Activates Amylin-3 Receptor Subtype by Triggering Multiple Intracellular Signaling Pathways. J. Biol. Chem. 2012; 287: 18820 - 18830.

Yasuto Yamaguchi, Xiao-Yan Du, Lei Zhao, John Morser, and Lawrence L. K. Leung. Proteolytic Cleavage of Chemerin Protein Is Necessary for Activation to the Active Form, Chem157S, Which Functions as a Signaling Molecule in Glioblastoma. J. Biol. Chem. 2011; 286: 39510 - 39519.

Yohei Okubo, Hiroshi Sekiya, Shigeyuki Namiki, Hirokazu Sakamoto, Sho Iinuma, Miwako Yamasaki, Masahiko Watanabe, Kenzo Hirose, and Masamitsu Iino. Imaging extrasynaptic glutamate dynamics in the brain. PNAS 2010; 107: 6526 - 6531.

†Prices are based on the websites of Sigma-Aldrich and Invitrogen as of April 2013 respectively, which may vary for different customers.

Ca2+ Indicator AAT Bioquest Sigma† Invitrogen†

Cal-520™ AM $195/10x50 µg not available not available

Fluo-8® AM $175/10x50 µg not available not available

Fluo-4 AM not available not available $218/10x50 µg

Fluo-3 AM $145/mg $390/mg $242/mg

Compare to Save

Pay Less. Perform Better.


Cat. # Product Description Size12530 Amplite™ Gaussia Luciferase Reporter Gene Assay Kit 1 plate

12518 Amplite™ Luciferase Reporter Gene Assay Kit *Bright Glow* 1 plate

12535 Amplite™ Renilla Luciferase Reporter Gene Assay Kit 1 plate

12500 Luciferase *Recombinant firefly* 1 mL

Gaussia Luciferase Reporter Gene Assay

Reporter Gene Analysis

The most versatile reporter gene is the firefly luciferase. Recently there is steadily increasing use of other luciferases, such as Gaussia luciferase since these reporters are smaller and do not require the presence of ATP. Gaussia luciferase is a 20 kD protein which catalyzes coelenterazine oxidation by oxygen to produce light. The bioluminescent enzyme derived from the marine copepod Gaussia princeps is efficiently secreted from mammalian cells upon expression.

AAT Bioquest's Amplite™ Gaussia Luciferase Reporter Gene Assay Kits use a proprietary luminogenic formulation to quantify luciferase activity in cell medium. The formulation generates a luminescent product that gives strong luminescence upon interaction with Gaussia luciferase. The kits provide all the es-sential components that are compatible with HTS liquid handling instruments. They have high sensitivity and can be performed in a convenient 96-well and 384-well microtiter-plate format. The “glow-type” signal with a half-life of one hour provides a consistent signal across large number of assay plates. The assay is compatible with standard cell growth media.

Figure 3.3. Secreted Gaussia luciferase culture medium was measured with Amplite™ Gaussia Luciferase Reporter Gene Assay Kit (blue line) and a commercially available Gaussia Luciferase Assay Kit (red line) respectively in a 96-well white plate with a NOVOstar plate reader (BMG Labtech).

Firefly Luciferase Reporter Gene Assay

The most versatile and common reporter gene is the luciferase of the North American firefly photinus pyralis. The protein requires no posttranslational modification for enzyme activity. It is not even toxic in high concentration (in vivo) and can be used in pro- and eukaryotic cells.

AAT Bioquest's Amplite™ Luciferase Reporter Gene Assay Kits use a proprietary DTT-free formulation to quantify luciferase activity in live cells and cell extracts. The assay is based on firefly luciferase, a monomeric 61 kD enzyme that catalyses a two-step oxidation of luciferin, which yields light at 560 nm. Our formulation generates a luminescent product that gives strong luminescence upon interac-tion with luciferase. The kits provide all the essential components with an optimized “mix and read” assay protocol that is compatible with HTS liquid handling instruments. They have high sensitivity and can be used for the assays that require low detection limit. The kits have a fast, simple, and homogeneous bioluminescence assay for studying gene regulation and function. The assay is compatible with the use of standard cell growth media.

Figure 3.4. Luciferase dose responses were measured with Amplite™ Luciferase Reporter Gene Assay Kit. The kit can detect as low as 0.1 pg/well luciferase with 20 minute to 5 hour incubation without losing signal intensity. The integration time is 1 second. The half life is more than 4 hours.

Cell-Based Assays www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304



www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304 Cell-Based Assays

iFluor™ 594, a New Bright Fluorescent Labeling Dyean Excellent Replacement to Texas Red®, DyLight™ 594 and Alexa Fluor® 594 Dyes

iFluor™ 594 has spectral characteristics similar to those of Texas Red®, DyLight™ 594 and Alexa Fluor® 594 with excitation and emission wavelength at ~592/614 nm when conjugated to pro-teins. iFluor™ 594 dye has superior labeling performance and better stability than Texas Red®. Our iFluor™ 594 conjugated streptavidin provides high fluorescence intensity and low background. Biomol-ecules conjugated to iFluor™ 594 exhibit little spectral overlap with green-fluorescent conjugates and can be efficiently excited by 568 nm line of Ar-Kr laser and by the 594 nm line of orange He-Ne laser. The minimal spectral overlap makes iFluor™ 594 an ideal second color in combination with a green color such as GFP, FITC, Alexa Fluor® 488 or iFluor™ 488. Our in-house research indicated that the iFluor™ 594-RPE conjugates demonstrate better FRET than Alexa Fluor® 594-RPE.

Ex (nm) 586

Em (nm) 611

EC (cm-1M-1) 120,000

CF280 nm* 0.0605

CF260 nm** 0.0945

* CF280 nm = EC280 nm/ECdye max (Correction factor for peptides and proteins) ** CF260 nm = EC260 nm/ECdye max (Correction factor for oligos and nucleic acids)

Quick Summary

Figure 3.5. The excitation and emission spectra of iFluor™ 594 Goat Anti-Rabbit IgG Conjugate (Cat# 16628) in PBS buffer (pH 7.2).

Figure 3.6. HeLa cells were stained with Rabbit HDAC antibody, and followed with iFluor™ 594 Goat Anti-Rabbit IgG Conjugate and Alexa Fluor® 594 Goat Anti-Rabbit IgG Conjugate respectively under the same conditions. The iFluorTM 594 Goat Anti-Rabbit IgG conjugate (left panel) demonstrated much lower staining background than the corresponding Alexa Fluor® 594 (right panel).

HD

AC

Ant

ibod

yN

egat

ive

Cont

rol

iFluor™ 594 Alexa Fluor® 594

Figure 3.7. HeLa cells were stained with mouse anti-tubulin followed with iFluor™ 594 Goat Anti-Mouse IgG (red, Cat# 16468), and nuclei were stained with Hoechst 33342 (blue, Cat# 17530).


Cat. # Product Description Alternative to Size

20073 Annexin V-iFluor™ 594 Conjugate Texas Red®, Alexa Fluor® 594, DyLight™ 594 100 tests

16468 iFluor™ 594 Goat Anti-Mouse IgG (H+L) Texas Red®, Alexa Fluor® 594, DyLight™ 594 200 µg

16628 iFluor™ 594 Goat Anti-Rabbit IgG (H+L) Texas Red®, Alexa Fluor® 594, DyLight™ 594 200 µg

2577 iFluor™ 594-RPE Tandem Texas Red®, Alexa Fluor® 594, DyLight™ 594 1 mg

16962 iFluor™ 594-Streptavidin Conjugate Texas Red®, Alexa Fluor® 594, DyLight™ 594 200 µg

1029 iFluor™ 594 Succinimidyl Ester Texas Red®, Alexa Fluor® 594, DyLight™ 594 1 mg

1230 ReadiLink™ iFluor™ 594 Protein Labeling Kit Texas Red®, Alexa Fluor® 594, DyLight™ 594 1 kit

Cell-Based Assays www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304

trFluor™ Fluorescent Labeling Dyes and Their Bioconjugates

*Optimized for TR-FRET Applications*

Many biological compounds present in cells, serum or other biological fluids are naturally fluorescent, and thus the use of conventional, prompt fluorophores leads to serious limitations in assay sensitivity due to the high background caused by the auto-fluorescence of the biological molecules to be assayed. The use of long-lived fluorophores combined with time-resolved detection (a delay between excitation and emission detection) minimizes prompt fluorescence interferences.

AAT Bioquest's trFluor™ probes enable time-resolved fluorometry (TRF) for the assays that require high sensitivity. These trFluor™ probes have large Stokes shifts and extremely long emission half-lives when compared to traditional fluorophores such as Alexa Fluor® or cyanine dyes. Compared to other TRF compounds, our trFluor™ probes have relatively high stability, high emission yield and ability to be linked to biomolecules. Moreover, our trFluor™ Eu probes are insensitive to fluorescence quenching when conjugated to biological polymers such as antibodies.

Key Features of trFluor™ Dyes:

• No fluoride addition is required.

• No enhancing solution is required.

• Available in a variety of reactive forms.

• Much easier to be conjugated to biomolecules.

• Much higher conjugation yield than other TRF dyes.

• Maximally excited by the common light sources at ~350 nm.

• trFluor™ Eu dye is optimized to pair with APC, iFluor™ 647, TF5, Cy5®, DyLight™ 650 and Alexa Fluor® 647.

• trFluor™ Tb dye is optimized to pair with FITC, iFluor™ 488, TF2, DyLight™ 488 and Alexa Fluor® 488.

trFluor™ Donors Recommended AcceptorstrFluor™ Eu iFluor™ 647, TF5, APC

trFluor™ Tb iFluor™ 488, FITC, TF2

Table 4.1. Typical acceptors for the time-resolved luminescent probes

trFluor™ Eu-Labeled Anti-IgG

TR-FRET

Emission @665 nmTF5

Emission @615 nmExcitation

Eu

Antibody Antigen

trFluor™ Dyes for TR-FRET Assays

Figure 4.1. TR-FRET assay principle using trFluorTM Eu as the donor while Tide FluorTM 5 (TF5) as the acceptor

www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304 Labeling Probes



Ex (nm) 346

Em (nm) 617

EC (cm-1M-1) ~22,000

CF280 nm* 0.777

CF260 nm** 0.911

* CF280 nm = EC280 nm/ECdye max (Correction factor for peptides and proteins); ** CF260 nm = EC260 nm/ECdye max (Correction factor for oligos and nucleic acids)

Quick Summary

trFluor™ Eu TRF Labeling Dye

AAT Bioquest's trFluor™ Eu probes enable TRF for the assays that require high sensitivity. The trFluor™ Eu dye has large Stokes shifts and extremely long emission half-lives when compared to more traditional fluorophores such as Alexa Fluor® or cyanine dyes. Compared to other time-resolved fluorescent probes, our trFluor™ Eu probes have relatively high stability, high emission yield and ability to be linked to biomolecules with higher conjugation yield. Moreover, our trFluor™ Eu probes are insensitive to fluorescence quenching when conjugated to biological polymers such as antibodies. To maximize its TR-FRET potential, trFluor™ Eu dye is optimized to pair with APC, iFluor™ 647, TF5, Cy5®, DyLight™ 650 and Alexa Fluor® 647.

trFluor™ Tb TRF Labeling Dye

AAT Bioquest's trFluor™ Tb dye has large Stokes shifts and extreme-ly long emission half-lives when compared to more traditional fluo-rophores such as Alexa Fluor® or cyanine dyes. Compared to other time-resolved fluorescent probes, our trFluor™ Tb probes have relatively high stability, high emission yield and ability to be linked to biomolecules with higher conjugation yield. Moreover, our trFluor™ Tb probes are insensitive to fluorescence quenching when conjugated to biological polymers such as antibodies. To maximize its TR-FRET potential, trFluor™ Tb dye is optimized to pair with FITC, iFluor™ 488, TF2, DyLight™ 488 and Alexa Fluor® 488.

Ex (nm) 330

Em (nm) 544

EC (cm-1M-1) ~20,000

CF280 nm* 0.797

CF260 nm** 0.942

Quick Summary

Figure 4.3. The excitation and emission spectra of trFluor™ Tb Goat Anti-Rabbit IgG conjugate (Cat# 16669) in PBS buffer (pH 7.2).

Figure 4.2. The excitation and emission spectra of trFluor™ Eu Goat Anti-Rabbit IgG conjugate (Cat# 16668) in PBS buffer (pH 7.2).

* CF280 nm = EC280 nm/ECdye max (Correction factor for peptides and proteins); ** CF260 nm = EC260 nm/ECdye max (Correction factor for oligos and nucleic acids)

ReadiLink™ iFluor™ Antibody Labeling Kits

ReadiLink™ iFluor™ Antibody Labeling Kits provide a convenient way to label antibodies using a stable reactive form of the iFluor™ dyes. The reactive iFluor™ dyes show good reactivity and selectivity with the aliphatic amines of antibod-ies and forms a carboxamide bond, which is identical to, and is as stable as the natural peptide bond. iFluor™-antibody conjugates may be used for immuno-fluorescent staining, fluorescent in situ hybridization, flow cytometry and other biological applications. Each kit comes with all the essential components for performing the conjugation reaction and for purifying the iFluor™-antibody conjugates. ReadiLink™ Kits only require two simple mixing steps to produce the desired conjugates for flow cytometry and fluorescence imaging applica-tions. The conjugation kits provide the best method for readily labeling small amount of antibodies without requiring column purification.

Key Features of ReadiLink™ Kits :

• Complete, all the components provided in the kits.

• Robust, only two simple mixing steps required.

• Rapid, less than 10 minutes hands-on time.



www.aatbio.com • [email protected] Tel: 800-990-8053 • Fax: 408-733-1304 Labeling ProbesLabeing Probes www.aatbio.com • [email protected]

Tel: 800-990-8053 • Fax: 408-733-1304


Cat. # Product Description Size Ex (nm) Em (nm)1300 ReadiLink™ trFluor™ Eu Protein Labeling Kit 2 labelings 346 617

1305 ReadiLink™ trFluor™ Tb Protein Labeling Kit 2 labelings 330 544

16518 trFluor™ Eu Goat Anti-Mouse IgG (H+L) 100 μg 346 617

16668 trFluor™ Eu Goat Anti-Rabbit IgG (H+L) 100 μg 346 617

1434 trFluor™ Eu Maleimide 100 μg 346 617

16925 trFluor™ Eu-Streptavidin Conjugate 100 μg 346 617

1433 trFluor™ Eu Succinimidyl Ester 1 mg 346 617

16519 trFluor™ Tb Goat Anti-Mouse IgG (H+L) 100 μg 330 544

16669 trFluor™ Tb Goat Anti-Rabbit IgG (H+L) 100 μg 330 544

1444 trFluor™ Tb Maleimide 100 μg 330 544

16926 trFluor™ Tb-Streptavidin Conjugate 100 μg 330 544

1443 trFluor™ Tb Succinimidyl Ester 1 mg 330 544


Cat. # Product Description Size Ex (nm) Em (nm)

1220 ReadiLink™ iFluor™ 350 Antibody Labeling Kit 2 labelings 345 442










Figure 4.4. ReadiLink™ Kit Labeling Principle • Step 1. Start the labeling reaction by mixing a labeling dye with a protein (to be labeled) in the Reaction Buffer (pH 7.5-8.5).• Step 2. Incubate the reaction solution and get a mixture of the desired protein conjugate and unreactive free dye. • Step 3. Quench the reaction by mixing a non-fluorescent Tide Quencher™ (TQ) dye with the reaction solution. The TQ dye stops the reaction

AND converts the unreactive free labeling dye to the non-fluorescent TQ-labeling dye complex, which eliminates the background fluorescence interference of the free labeling dye.

Austria:Biomol GmbHEmail: [email protected]: http://www.biomol.de

Australia:Life Research Pty Ltd.Email: [email protected]: http://www.liferesearch.com

Belgium:Gentaur BVBAEmail: [email protected]: http://www.gentaur.com

Canada:Cedarlane Laboratories Ltd.Email: [email protected]: http://www.cedarlanelabs.com

China:AmyJet Scientific Inc.Email: [email protected]: http://www.amyjet.com

Beijing Zhonghao Shidai Co., Ltd Email: [email protected]: http://www.biopcr.com

Mai Bio Co., LtdEmail: [email protected]: http://www.maibio.com

Tianjin Biolite Biotech Co., Ltd Email: [email protected]: http://www.tjbiolite.com

Croatia:Biomol GmbHEmail: [email protected]: http://www.biomol.de

Czech Republic:Scintila, s.r.o.Email: [email protected]: http://www.scintila.cz

Denmark:Nordic BioSite ApSEmail: [email protected]: http://www.nordicbiosite.dk

Estonia:Biomol GmbHEmail: [email protected]: http://www.biomol.de

Nordic BioSite ABEmail: [email protected]: http://www.biosite.se

Finland:Nordic BioSite OYEmail: [email protected]: http://www.biosite.fi

France:EUROMEDEXEmail: [email protected]: http://www.euromedex.com

InterchimEmail: [email protected]: http://www.interchim.com

Germany:Biomol GmbHEmail: [email protected]: http://www.biomol.de

Hong Kong:Mai Bio Co., LtdEmail: [email protected]: http://www.maibio.com

Hungary:IZINTA Trading Co., Ltd.Email: [email protected]: http://www.izinta.hu

Iceland:Nordic BioSite ABEmail: [email protected]: http://www.biosite.se

India:Biochem Life SciencesEmail: [email protected]: http://www.bcls.in

GenxBio Health Sciences Pvt. Ltd,Email: [email protected]: [email protected]: http://www.genxbio.com

Ireland:Stratech Scientific Ltd.Email: [email protected]: http://www.stratech.co.uk

Israel:ADVANSYS Technologies for Life Ltd.Email: [email protected]: http://www.advansys.co.il

Italy:Space Import Export S.r.l.Email: [email protected]: http://www.spacesrl.com

Valter Occhiena S.r.l.Email: [email protected]: http://www.valterocchiena.com

Japan:Cosmo Bio Co., Ltd.Email: [email protected]: http://www.cosmobio.co.jp

Nacalai Tesque, Inc.Email: [email protected]: http://www.nacalai.com

Wako Pure Chemical Industries, Ltd.Email: [email protected]: http://www.wako-chem.co.jp

Korea:Cheong Myung Science CorporationEmail: [email protected]: http://www.cmscorp.co.kr

Latvia and Lithuania:Nordic BioSite ABEmail: [email protected]: http://www.biosite.se

Netherlands:EUROMEDEXEmail: [email protected]: http://www.euromedex.com

Norway:Nordic BioSite ABEmail: [email protected]: http://www.biosite.se

Poland and Slovenia:Biomol GmbHEmail: [email protected]: http://www.biomol.de

Romania:SC VitroBioChem SRLEmail: [email protected]: http://www.vitro.ro

Singapore and Other South Asian Countries:BST Scientific Pte Ltd.Email: [email protected]: http://www.bstsci.com

Slovakia:Scintila, s.r.o. Email: [email protected]: http://www.scintila.cz

South American Countries and Regions:Impex Comércio Internacional Ltda.Email: [email protected]: http://www.impexbrasil.com.br

Spain and Portugal:Deltaclon S. L Email: [email protected]: http://www.deltaclon.com

Sweden:Nordic BioSite ABEmail: [email protected]: http://www.biosite.se

Switzerland:LuBioScience GmbHEmail: [email protected]: http://www.lubio.ch

Taiwan:Rainbow Biotechnology Co., LTD.Email: [email protected]: http://www.rainbowbiotech.com.tw

Turkey:Biomol GmbHEmail: [email protected]: http://www.biomol.de

United Kingdom:Stratech Scientific Ltd.Email: [email protected]: http://www.stratech.co.uk

International Distributors

AAT Bioquest®

Advancing Assay & Test Technologies

Documents

oue ssue Assayise Letters - AAT Bioquest · 2/1/2013 · atiometrR uri F a Caium ndiaI orst 1 onadioatie uose ptake Assays ... all products are for Research Use Only. ... 21050 Quin-2