Visual Protein 2016 Catalog

for Protein Science

Table of Contents

Western Blotting

Protein Purification

Antibody Production

Proteomics

LuminolPenTM HRP System LuminolPenTM Lite HRP SystemBlockPROTM Blocking BufferBlockPROTM Protein-Free Blocking BufferLumiFlash™ Prime Chemiluminescent SubstrateLumiFlash™ Ultima Chemiluminescent SubstrateLumiFlash™ Infinity Chemiluminescent Substrate

ImmunoFastTM AdjuvantHybriMore™ Hybridoma Cloning Factor

ExtracPROTM Protein Extraction ReagentFracPROTM Protein Fractionation KitPhosPROTM Phosphoprotein Purification Series

VisPROTM 5 Minutes Protein Stain KitIEF Optimizer

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1012

141617

2023

Western blotting (WB) is the most utilized technique in current life science research. How-ever, due to the complexities of biological materials and the insufficient specificity of some antibodies, intriguing WB results are generally observed, such as “multiple band signals” or “hybridized bands” coming with unsure molecular weight. To validate a WB result, calculat-ing the molecular weight of the obtained protein signals may be the most straightforward meathod. Nevertheless, since most pre-stained protein standard markers can’t be visual-ized after the hybridization procedure, many researchers may align the WB result (cap-tured image) with the original blot membrane. Such procedure is tedious and may intro-duce estimation bias.

LuminolPen™ Series is a perfect tool for chemiluminescent detection in Western blot. With simply writing on the pre-stained marker, the position of the molecular weight can be showed on the image. It could also help you to note the experiment condition or evaluate the efficacy of ECL substrate. We offer two versions of LuminolPen based on their chmilu-minesent properties: enzyme-based (LH03) and chemical based (LH604).

How does LuminolPen improve the correctness in distiquishing target signals?

Distinguish target signal and non-specific binding signal with LuminolPen. 30 µg cell lysate SP-1 and detect with anti-ADNP (mouse, 1:2,000). Secondary antibody : anti-mouse IgG-HRP (1:15,000). Membrane : Hyond™ P. Detection : Hyperfilm™ ECL. Signal exposed for 30 seconds and detected by X-ray. The molecular weight note by LuminolPen™ reveal the correct target signal (ADNP 124KDa) on X-ray film.

How does LuminolPen tell the efficacy of ECL substrates?

Evaluate ECL substrate efficacy by LuminolPen™. 30 µg cell lysate HepaG2 and detect with anti-AMP-active protein kinase (mouse, 1:1,000). Secondary antibody : anti-mouse IgG-HRP (1:10,000). Membrane : Hyond™ P. Detection : Hyperfilm™ ECL. Signal exposed for 30 seconds and detected by X-ray. Signal result present by normal ECL substrate is stronger than using expired ECL substrate. LuminolPen™ noted marker can be an indicator to check the result or adjust different experiment to similar experimental condition.

LuminolPenTM Series

2

Performance

Labeling molecular weight and note on the membrane with LuminolPen™, HRP System.

LuminolPen™ HRP System is the world’s first luminescent color development marker pen.The enzyme-based property makes it suitable for most brands of ECL substrate.

LuminolPenTM HRP System

Ordering Information

LuminolPen™ HRP SystemLH03-50 1 penAbout 1,000 membranes drawing

LH03-10 1 penLuminolPen™ HRP SystemAbout 100 membranes drawing

LH603

3

Unique - the world’s first luminescent color development marker pen

Compatible - applicable with most brands of ECL substrate Convenient - able to make note and show on the image with the data resultHelp you to evaluate the efficacy of your ECL substrate

LuminolPen™ Lite HRP System provides easy writing double heads.

LuminolPen™ Lite HRP System is second generation prod-uct of LuminolPen™ which is also designed for chemilumi-nescent detection in Western blot. Lh604-500/Lh604-200 is designed for drawing a blot which will be expososed in 10 mins. LH604-500L/LH604-200L is for using when longer exposure time is required (10 min to 1 hour).

粗筆頭

細筆頭 Heavy head

Fine head

LuminolPenTM Lite HRP SystemLH604

Performance

Ordering Information

LH604-200 1 penAbout 500 membranes drawing

LH604-30 1 penAbout 100 membranes drawing

LuminolPen™ Lite HRP SystemLH604-200L 1 penAbout 500 membranes drawing

LH604-30L 1 penLuminolPen™ Lite HRP SystemAbout 100 membranes drawing

4

Lower price - with smaller package than LuminolPen™ and lower cost than ECL

More Convenient - just writing before image captureMore Choice - with different exposure time and volume to be choose

Western blotting marker

LuminolPen™ Lite HRP System

LuminolPen™ Lite HRP System

BlockPROTM Blocking Buffer

BlockPROTM Blocking Buffer is suitable for blocking in Western blot, ELISA, immunohistochemistry and other immunochemical application. It can block excess bind-ing site but not cover on the binding protein and there-fore increase the signal intensity. Present better data result than milk and BSA blocking buffer.

Ordering Information

BlockPROTM Blocking BufferBP01-1L 1 kit

Skim Milk

AMPK

Casein BSASkim Milk

Performance

Signal strength comparison of Casein, BSA, milk, and BlockPRO™ Blocking Buffer. Loading 30 µg cell lysate (HepaG2) and detect with anti-AMPK (mouse, 1:1,000). Secondary antibody : anti-mouse IgG-HRP 1:10,000. Membrane: Hyond™ P. Detection : Hyperfilm™ ECL. All results were exposed for 30 seconds and capture by X-ray film.

Blocking with skim milk or BlockPRO™ Blocking Buffer and detect by SEM. 30 µg cell lysate (HepaG2) separated by 12.5% SDS-PAGE and blocking by skim milk or Block-PRO™ Blocking Buffer. Membranes detected by SEM. The SEM result showed that BlockPRO™ Blocking Buffer only blocking on excess binding site and reveal the position of antigen and therefore will not reduce the signal intensity.

Effective Blocking

Better Signal Intensity than Traditional Blocking Agents

500mL x 2

BP01

5

Ordering InformationBlockPROTM Protein-Free Blocking Buffer

BlockPROTM Protein-Free Blocking Buffer (10X)

BlockPROTM Protein-Free Blocking Buffer (10X)

BF01-1L 1 kit500mL x 2

BF10-100 1 kit100mL x 1

BF10-200 1 kit100mL x 2

Performance

BlockPROTM Protein-Free Blocking Buffer

BlockPROTM Protein-Free Blocking Buffer is a non-protein formulation which enhances sensitivity and minimizes background noise, presenting better results than tradi-tional protein-based blocking buffer in immunoassays. The synthetic formulation of BlockPROTM Protein-Free Blocking Buffer makes it suitable for PVDF and nitrocellu-lose platform, avidin/biotin system, detection of phospho-protein, and other immunochemical applications.

Better Sensitivity & Less Background Noise

Sutible For Multiple Protein Antigens

BlockPROTM Protein-Free Blocking Buffer is better than protein-based blocking buffers (skim milk, BSA and casein) for detection of target protein in Western blotting

Sample: THP-1 cell lysatesBlocking time: 1 hourTarget antigen: pAMPKExposure time: 30 sec

BlockPROTM Protein-Free Blocking Buffer is suitable for different kinds of protein detection.

Sample: THP-1 cell lysatesBlocking time: 1 hourExposure time: 30 sec

5% skim milk

3% BSA

2% casein

BlockPROTM Protein-Free Blocking Buffer

THP-1 cell lysate (µg protein)

15 7.5 3.75 1.88 0.94 0.47 0.23

BF01

6

LumiFlash™ Prime Chemiluminescent Substrate is a ready-to-use reagent for chemiluminescent detection of immobilized proteins (Western blotting), conjugated with horseradish peroxidase (HRP) directly or indirectly. In the presence of hydrogen peroxide, HRP catalyzes the oxidation of cyclic diacylhydrazides, such as luminol, and light emits. LumiFlash™ Prime Chemiluminescent Substrate provides a convenient way to visualize HRP-based detection. Simply mix and add the solutions onto the membrane. The signal of target protein can be recorded by exposure to X-ray film or compatible image acquisition system.

Economic and stable ECL substrate for Western blotting

The comparison of signal intensity of LumiFlash Prime, VisGlow and Luminata Crescendo on Western blotting. Hela cell lysate with 1/2 serial dilution from 20 µg was separated by 12.5% SDS-PAGE and probed by anti b-actin. All results were exposed to X-ray film for 1 minute.

Performance

High sensitivity - detect protein target to picogram level

Antibody saving - less antibody usage with higher quality result on Western blotting Economic - offer excellent quality with lower price Low background - provide low background on the WB result

LumiFlashTM Prime Chemiluminescent Substrate

Ordering InformationLF01-100 1 kit

50mL soultion A + 50mL solution B

and ELISA application

LF01LumiFlash™ Prime Chemiluminescent Substrate

7

LumiFlashTM Prime Chemiluminescent SubstrateLF01-500 1 kit250mL soultion A + 250mL solution B

LumiFlash™ Ultima Chemiluminescent Substrate, HRP is an advanced ECL product in terms of efficacy and sensitivity for chemiluminescent detection of immobilized proteins (Western blotting and ELISA). LumiFlash™ Ultima Chemiluminescent Substrate, HRP provides high sensitivity and long signal duration in Western blotting application. It needs very short exposure time to the X-ray film or other documentation systems, and can get low background and high signal with clear image. LumiFlash™ Ultima Chemiluminescent Substrate, HRP is the perfect choice for most Western blotting application.

High signal sensitivity and long signal duration

The comparison of signal intensity of LumiFlash Ultima, VisGlow Plus and Luminata Crescendo on Western blotting. K562 cell lysate with 1/2 serial dilution from 50 µg was separated by 12.5% SDS-PAGE and probed by anti HSP 47. All results were exposed to X-ray film for 3 minutes.

Performance

High sensitivity - detect protein target to low-picogram level

Antibody saving - less antibody usage with higher quality result on Western blotting Economic - cost less than other ECL substrate with similar sensitivity level Long duration - provide stable signal for long exposure

LumiFlashTM Ultima Chemiluminescent Substrate

Ordering InformationLF08-100 1 kit

50mL soultion A + 50mL solution B

and ELISA application

LF08LumiFlash™ Ultima Chemiluminescent Substrate

8

LumiFlashTM Ultima Chemiluminescent SubstrateLF08-500 1 kit250mL soultion A + 250mL solution B

LumiFlash™ Infinity Chemiluminescent Substrate, HRP is an extremely sensitive enhanced chemiluminescent substrate for femtogram-level by detection of immobilized proteins (Western blotting). In Western blotting or ELISA application, LumiFlash™ Infinity Chemiluminescent Substrate, HRP provides high signal and low background, which allows detection of target protein on PVDF or nitrocellulose. This feature benefits the researchers with excellent low background result and without signal burn effect at the same time.

A highly sensitive ECL substrate for Western blotting analysis

The comparison of signal intensity of LumiFlash Infinity and Luminate Forte on Western blotting. Hela cell lysate with 1/2 serial dilution from 20 µg was separated by 12.5% SDS-PAGE and probed by anti AMPKa1. All results were exposed to X-ray film for 5 minutes.

Performance

High sensitivity - detect protein target to femtogram level

Antibody saving -

Low background -

LumiFlashTM Infinity Chemiluminescent Substrate

Ordering InformationLF16-100 1 kit

250mL soultion A + 250mL solution B

use very low antibody concentration on the WB and ELISA applica-tion; dilute primary antibody 5000 to 20,000-fold and secondary antibody 100,000 to 400,000-fold (from 1mg/mL stock)

provide insignificant background and avoid non-specific signals on the WB result

LF16LumiFlash™ Infinity Chemiluminescent Substrate

9

LumiFlashTM Infinity Chemiluminescent SubstrateLF16-500 1 kit250mL soultion A + 250mL solution B

Strong - elicit strong immunorespond with less antigen loading

Performance

Adjuvants can be categorized into Antigen delivery system and Immunoptentiator. Antigen delivery system works by the adjuvant surrounding the antigen and can be easily recognized by macrophage cells. Immunopotentiator is an adjuvant with Toll-Like Receptor (TLR) which induces the immune cells to enhance immune-response. Such are MPI and MPD.

ImmunoFast™ Adjuvant is a new aqueous emulsify adjuvant which can efficiently elicit significant immunorespond and induce high amount of IgG production in very short time. The aqueous texture makes antigen mixture and injection much easier and the special designed non toxicity formula provide higher survival rate of immunize animal during experiment period.

ImmunoFastTM can elicit much stronger immune-response in animals than other adjuvants

After performing a regularimmunization procedure on mice, Immuno-Fast delivers approximate 10 fold stronger Ab titer than CFA (Complete Freud’s adjuvant) and IFA (Incomplete Freud’s adjuvant). ImmunoFast may also shorten the period of immunization with sufficient Ab titer.

Simple - easy mixture characteristic with the aqueous textureSafe - non toxicity and harmful effect to immunize animalEffective - high efficiency of antibody production and induce complete class switch

ImmunoFastTM AdjuvantIF01

10

from IgM to IgG

High efficiency of antibody production

The comparison of immunoresponse elicited by ImmunoFast and CFA at the 2nd and the 6th week after first immunization. Mice were immunized with 50ug of bovine serum albumin in Immuno-FastTM & CFA (complete Freund Adjuvant). The adjuvant boosted in 2nd, 4th and 6th week. Then serum IgG tilters were measured by ELISA.

For 4 injections for mouse

ImmunoFastTM can induce high amount of IgG production

Levels of the elicited immunoglobulin molecules in an ImmunoFast boosted animal. Mice were immunized with 50ug of bovine serum albumin in ImmunoFastTM. The adjuvant boosted in 2nd, 4th and 6th week. Then IgG and IgM tilters were measured by ELISA.

For 20 injections for mouse

Ordering InformationImmunoFast™ Adjuvant

ImmunoFast™ AdjuvantIF01-20N

IF01-4N

11

Performance

HybriMore™ Hybridoma cloning factor is a special supplement which is adding in the medium when culturing hybridoma cells. It can substantially provide necessary growth factor during cell culture and therefore successfully increase the cloning efficiency and raise the survival rate of hybridoma cell. No negative effect to hybridoma cell with defined chemical component and defined concentration.

Up to 10 fold increasing numbers of recovered hybridoma clones

The newly PEG fused hybridoma cells were plated into a 96-well plate containing FCS media with HybriMore (green bars), FCS media with feeder layer (blue bars), or regular FCS media (white bars). Hybridoma cells were subject to HAT selection 14 days after the cell fusion. Two mouse myeloma fusion partners, NS-1 and SP2/0, were evaluated by four independent fusion experi-ments with freshly prepared mouse spleens. A significant higher cloning efficiency was observed for the usage of HybriMore.

Increase cloning efficiency and cell survival rateCulturing hybridoma cells with better antibody secretion and productionGrowth promoting supplement with defined chemical component and concentration

Increase the successful rate of monoclonization during monoclonization

Eight clones of hybridoma cells from NS-1 or SP2/0 fusion partners were monoclonized in the media containing FCS media with HybriMore (green bars), FCS media with feeder layer (blue bars), or regular FCS media (white bars). The numbers of viable hybridoma colonies in a well were visually counted under a microscope. A significant higher successful rate of monocloniza-tion was observed for the usage of HybriMore.

HB01HybriMore™ Hybridoma Cloning Factor

12

Ordering Information

Defined chemical, no animal source materials, no effect of cell physiology

HybriMoreTM, Hybridoma Cloning FactorHB01-1L 1 bottle

HybriMore

Optimal numbers of hybridoma clones

Defined chemical, noanimal source materials

No competition for nutrients

Defined concentration of supplement

Convenient to supplement

Feeder layer (conventional)

Overgrowth of newly formed hybridoma

Source of contamination

Competition for nutrients

Variations in growth factor concentration

Tedious procedure to prepare the feeder layer

A clone of hybridoma cells (anti human transfer-rin, L3B5) was cultured in the media containing FCS media with HybriMore (green bars), FCS media with feeder layer (blue bars), or regular FCS media (white bars) for seven days. The superna-tants were harvest and examined by the titer of secreting Ab by ELSIA assay. The usage of Hybri-More will not alter the yield of secreting Ab in hybridoma cells.

Comparison of the application in hybridoma experiment

For 1 L culture medium dilution

13

Protein extraction is the starting step for sample preparation in most experiment. Protein can be extracted by a few methods such as detergent lysis, shearing force, treatment with low ionic salt (salting out). The biggist challenge in protein extraction is to generate a cellular extract that can be efficiently manipulated in downstream processes without inactivation or degradation of labile protein targets.

Yields of extracted protein are higher with ExtractPRO than freeze & thaw

Protein extract efficiency with different sample source. Cell line (K562), animal tissue (pig liver), fungi (PE1), and bacteria (E. coli) were extracted by freeze and thaw or ExtractPRO, analyzed with 12.5% SDS-PAGE, and stained by CBR method. The results indicate that ExtractPRO™ show better protein extract efficiency than traditional freeze and thaw method.

Performance

ExtractPROTM Protein Extraction Reagent is a new formula reagent for wide range of bio-samples protein extracting and can remove most of DNA, RNA, and cell debris with simple extraction procedure. ExtractPRO™ Protein Extraction Reagent can effectively collect protein sample without using protease inhibitor and the sample can also be preserved for long term after adding ExtractPRO™ reagent.

Shorten experiment time with lower costProtease inhibitor and freeze/thaw cycle are not required

Compatible for most bio-samples, including plant, animal, tissue, and microorganism protein extraction

Effectively maintain protein condition during long term preservation

EP05ExtracPROTM Protein Extraction Reagent

14

Ordering InformationExtractPROTM Protein Extraction ReagentEP05-30 1 bottle

30mL solution

Ensure high quality extracted proteins than freeze & thaw

Comparison of sample preparatrion by the ExtractPROTM Protein Extraction Reagent with freeze and thaw method. HepaG2 cell is prepared by traditional sample preparation (freeze & thaw) and ExtractPROTM Protein Extraction Reagent. The 2D-PAGE treated by ExtractPRO has lower background than by traditional sample preparation.

Total protein extract amount after storage in ExtractPRO. Cell line SP-2 preserved in Extract-PRO without protease inhibitor for 0, 14, and 28 days. Protein quantified by bradford method.

The protein portion in sample is well preserved during long-term storage in ExtractPRO

15

The FracPROTM Protein Fractionation Kit is a simple, reproducible and ultracentrifuge-independent method to separate four subcellular protein fractions, including nuclear, cytosol, membrane/organelle, and cytoskeletal fractions, in a detergent based procedure. The FracPROTM Protein Fractionation Kit takes advantage of the differential solubility of proteins in various subcellular compartments and utilizes highly specialized fractionation buffers to target specific subcellular compartments and simultaneously preserves the structural integrity of the proteins during each sequential fractionation.

Isolate four protein fractions with minimal cross-contamination

The fractionation efficacy of FracPRO™ Protein Fractionation Kit is presented by Western blot.

Sample: Human leukemia cell line (K562)(A) Heat shock protein 70 antibody (B) ATPase antibody (C) Histone deacetylase 2 antibody (D) Vimentin antibody.

Performance

Ordering Information

FracPROTM Protein Fractionation KitFP01-5 1 kit10mL Cytosol buffer 10mL Membrane/Organelle buffer 7.5mL Nuclear buffer 1.5mL Cytoskeletal buffer

100mL Cytosol buffer 100mL Membrane/Organelle buffer 75ml Nuclear buffer 15ml Cytoskeletal buffer

FracPROTM Protein Fractionation KitFP01-50 1 kit

55 kDa

100 kDa

55 kDa

55 kDa

Marker

Cytosol Fr

action

Membran

e Frac

tion

Nuclear

Fractio

n

Cytoske

letal F

ractio

n

A. ALDH2 (56.4 kDa)

B. ATPase (112 kDa)

D. Vimentin (57 kDa)

C. HDAC2 (55.4kDa)

Variable sample application including cultured cells and mammalian tissuesSeparate and collect four subcellular protein fractions in short timeIsolate high quality protein fractions with easy procedure

FracPROTM Protein Fractionation KitFP01

16

PhosPROTM Phosphoprotein Purification Series

The investigation of protein phosphorylation is of key interest in current proteomics. It has been estimated that phosphoprotein accounts for 10-20% of total cellular proteins. PhosPROTM Phosphoprotein Purification Series are designed to enrich phosphorylated proteins up to 5-10 folds, and thus facilitate the following detection of the minor phosphoproteins and the down-stream applications.

PhosPRO™ Series provides stringent buffer stock for customer to optimize washing condition for their sample and to maximize target phosphoprotein purification result

PP05PP03

Immobilized metal-affinity chromatography (IMAC)-based technique

Strategy of immobilized metal affinity chroma-tography (IMAC).

PhosPRO™ Series has 2 versions with different packages for specific purposes

PhosPROTM Phosphoprotein Purificaction Series is based on immobilized metal affinity chromatography (IMAC). It is thought that the phosphate groups on proteins or peptides interact with the metal ions immo-bilized to the chelating ligand, such as iminodiacetic acid (IDA) or nitrilotriacetic acid (NTA) on the chromatographic media. Employing metal ion, such as ferric (Fe3+), gallium (Ga3+) or zinc (Zn3+) or other metal ions charged IMAC resin, numbers of studies have successfully enriched phosphoproteins or phosphopeptides from model proteins or biological samples .

Considering the complexity and diversity of proteomes from different biological materials, the purification results would vary for individual sample. PhosPROTM Phosphoprotein Purificaction Series provides a stringent buffer stock for optimizing the chromatographic washing condition. Users might make their own washing buffer for minimizing the binding of non-phosphopro-teins and maximizing the binding of phosphoprotein.

Optimized phosphopeptide enrichment protocol (next page)

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Performance

Comparison of the purification efficacy of PhosPROTM Phosphoprotein Enrichment Kit, PhosPROTM Phosphoprotein Purification kit, and Brand Q kit. Protein standard markers containing glycogen phosphorylase b (GP), bovine serum albumin (BSA), ovalbumin (OVA), carbonic anhydrase (CA), trypsin inhibitor (TI) and lactalbumin (LAC) were used for evaluation. Ovalbumin was the only phosphoprotein in the test material. F: flow through; E: eluent. VisPROTM 5 minutes Protein stain kit (VisPRO) and Pro-Q Diamond stain (Pro-Q, from phosphoproteins).

PhosPROTM Phosphoprotein Purificaction Series delivers excellent purification result for phosphoproteins. When a standard protein mixture was used as the testing material, PhosPROTM Phosphoprotein Purificaction Series specifically enriched the phosphoprotein, ovalbumin, from other nonphosphoproteins as shown in the figure below. Almost none of non-phosphoproteins are copurified in the elution fraction. With our proprietary chromatographic technology, Phos-PROTM Phosphoprotein Purificaction Series delivers excellent efficacy of phosphoprotein purification, as compared with other commercial kit.

Optimization condition protocol for PhosPRO Phosphoprotein purifi cation series.

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Ordering Information

Selection Tips

PhosPROTM Phosphoprotein Enrichment KitPP03-2C 1 kit2 pre-packed column 2mL Lysis buffer 250mL System buffer 100mL Elution buffer100mL Stringent buffer stock

PP03-6C 1 kitPhosPROTM Phosphoprotein Enrichment Kit6 pre-packed column 2mL Lysis buffer 250mL System buffer 100mL Elution buffer100mL Stringent buffer stock

PP03-5E 1 kitPhosPROTM Phosphoprotein Enrichment Kit3mL resin2mL Lysis buffer 250mL System buffer 100mL Elution buffer100mL Stringent buffer stock

PhosPROTM Phosphoprotein Purification KitPP05-2C 1 kitPre-packed cloumn package2 pre-packed column 2mL Lysis buffer 250mL System buffer 100mL Elution buffer100mL Stringent buffer stock

PP05-6C 1 kitPhosPROTM Phosphoprotein Purification KitPre-packed column package6 pre-packed column 2mL Lysis buffer 250mL System buffer 100mL Elution buffer100mL Stringent buffer stock

PP05-5E 1 kitPhosPROTM Phosphoprotein Purification KitEvaluation package 3ml resin 2mL Lysis buffer 250mL System buffer 100mL Elution buffer100mL Stringent buffer stock

PhosPROTM Phosphoprotein Enrichment Kit -Acheives higher yields of the phosphoproteins than other commercial kits

PhosPROTM Phosphoprotein Purification Kit -Acheives high purity of the isolated phosphoproteins

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Product highlights

Fast - quick staining procudureHigh sensitivity - detect protein level to 1 ngCompatibility - applicable with downstream procedure

The imidazole-zinc reverse stain, utilizing imidazole and zinc ions for protein visualization on electrophoretic gels, was originally introduced in the 1990s. This method is based on the selective precipitation of the imidazole-zinc complex in the gel, except in zones where proteins or other macromole-cules are present. Similar to negative film, this method only stains the gel rather than the target proteins. The VisPROTM

5 minutes Protein Stain Kit is a modified version of the imid-azole-zinc reverse stain, designed for convenience and superior performance.

Most protein gel staining methods are based on the principles of positive staining. To obtain an ideal result, the dye molecules must be completely absorbed by target proteins, and that may require a prolonged incubation of many hours, even up to one day. Furthermore, a tedious de-staining procedure may be required to remove the undesirable staining from the background.

Positive staining vs. Reverse staining

Unlike conventional positive staining methods, reverse staining can be completed in a short time because the target of staining is the gel rather than the proteins. Therefore, reverse staining methods allow researchers to obtain the results promptly.

VP01 VP05 VP10

VisPROTM 5 Minutes Protein Stain Kit

20

Comparison of staining sensitivity

Numerous tests have demonstrated that VisPROTM 5 Minutes Protein Stain Kit delivers better staining sensitivity than SYPRO RubyTM stain, silver stain, and Coomassie Brilliant Blue stain (CBR stain). The commercial protein markers (Amersham, GE healthcare) were used for evaluation. Numbers shown above the gels indicate the actual amount of bovine serum albumin protein (indicat-ed by arrow).

1155

ng

557

ng

288.

7 ng

144.

3 ng

72.2

ng

36.1

ng

18.0

ng

9.02

ng

4.52

ng

2.25

ng

1155

ng

557

ng

288.

7 ng

144.

3 ng

72.2

ng

36.1

ng

18.0

ng

9.02

ng

4.52

ng

2.25

ng

(A) VisPRO 5 minutes Protein Stain (B) Sypro Ruby Stain

(C) Silver Stain (D) CBR Stain

Performance

VisPROTM 5 Minutes Protein Stain kit perfectly matches to current proteomic research. More protein spots can be found on the two-dimensional electropho-resis (2-DE) gel developed with the VisPROTM 5 Minutes Protein Stain Kit (right) than the one developed with other high sensitivity protein staining methods, such as silver stain (left).

MALDI-TOF analysis of protein spots visualized with the VisPROTM 5 minutes Protein Stain Kit. The gel spot of rabbit phosphorylase b (10 ng) was cut and subjected to mass spectrometry. Twen-ty-two out of fifty signals were categorized as tryptic fragments of phosphorylase b.

VisPROTM 5 Minutes Protein Stain Kit perfectly matches to current proteomic research

21

Ordering Information

Method

Operate time

Sensitivity

Irritant or toxic chemical

Mass compatibility

Downsream application

VisPROTM 5 Minutes Protein Stain Kit

5~10 minutes

< 1ng

None

Good

1. Western Blotting2. Electric eluting

SYPRO Ruby

18 hrs

~1ng

Acetic acid, Methanol

Ok

No

Silver Stain

4~20 hrs

~1ng

Acetic acid, Silver Nitrate,

Glutaraldehyde

Bad

No

CBR Stain

1~8 hrs

50ng

Acetic acid, Methanol

Ok

No

Comparison of VisPROTM 5 Minutes Protein Stain Kit with other com-monly used methods

22

VisPROTM 5 Minute Protein Stain KitVP01-125 1 kit25mL solution 1 + 125mL solution 2

VisPROTM 5 Minute Protein Stain KitVP01-500 1 kit500mL solution 1 + 500mL solution 2

VisPROTM 5 Minute Protein Stain Kit (5X)VP05-125 1 kit500mL solution 1 + 500mL solution 2

VisPROTM 5 Minute Protein Stain Kit (5X)VP05-500 1 kit500mL solution 1 + 500mL solution 2

VisPROTM 5 Minute Protein Stain Kit (10X)VP10-1L 1 kit1000mL solution 1 + 1000mL solution 2

IEF Optimizer is a novel and convenient instrument which can measure the salt content of the two-dimensional electrophoresis (2-DE) samples. In combined with IEF salt meter and the online accessible voltage-hour calculator, IEF Optimizer can provide you a suggested total voltage hour for IEF process, avoiding underfocusing and overfocusing of proteins and not only give you reproducible data but also accuracy.

Providing the accurate salt content measurement of 2-DE samples and optimal IEF total voltage hour for 2-DE samplesIncreasing the reproducibility of 2-DE resultReducing underfocusing and overfocusing of 2-DE experiments, particularly analyz-ing precious clinical samples during DIGE experiments

Two-dimensional electrophoresis (2-DE) is widely used in proteomic researche. 2-DE combines first dimensional isoelectric focusing (IEF) to distinguish proteins according to their isoelectric point (pI), and second dimensional sodium dodecyl sulfate polyacryl-amide gel electrophoresis (SDS-PAGE) to subsequently separate proteins according to their molecular mass. Since being first introduced, numerous efforts have been made to improve the reproducibility of 2-DE. Salt interference in the first dimensional IEF is thought to be a major factor deteriorate the results of 2-DE. Salt contents in 2-DEsamples may result in high conductivity, and high heat, which burns the immobilized pH gradient (IPG) gel strips. Residual salt contents may decrease the progression of IEF; thereby worsening the results of protein focusing.

Product highlights

Salt interference make underfocusing and overfocusing in 2-DE gel

IEF OptimizerIOR01

23

Ordering Information

IEF Optimizer operation flow chart

IEF Optimizer providesyou the optimum parameters.Sample: pig liver, 2-D electropho-resis. 1-D electrophoresis is from pH 3-10. 2-DE using 12.5% SDS-PAGE. (LEFT) Results obtained according to the manufacturers’ instruction handbook. (RIGHT) Result obtained according to the IEF Optimizer parameter.

IEF OptimizerIOR01

IEF Salt Meter + Voltage-Hour Calculator (Online Software)

registration card

PerformanceOptimum parameters provided by IEF Optimizer show better results than the suggested protocol

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