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Buffers, Solutions, and Chemicals

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Zombie Aqua™/ Zombie NIR™ /Zombie Yellow™

Example Data:

PI and 7-AADZombie Aqua™/ Zombie NIR™ /Zombie Yellow™

Live Cell/Dead Cell DiscriminationIt is critical to understand the degree of cell death in any flow cytometry assay and exclude those cells from the analysis. BioLegend provides DNA dyes, Propidium Iodide and 7-AAD, that enter and stain dead cells, but are impermeable to live cells for rapid, cost-effective analysis of unfixed cells. In cases where cell fixation is required, we now introduce fixable Zombie Aqua™, Zombie NIR™, and Zombie Yellow™.

Excitation/Emission

One day old splenocytes were stained with Zombie Aqua™ and analyzed without fixation (blue) or analyzed after fixation and permeabilization (red). Cells alone without Zombie Aqua™ staining are indicated in black.

Excitable by the violet laser (405nm), Zombie Aqua™ emits into the Brilliant Violet 510™ channel, Zombie NIR™ emits into the APC-Cy7 channel, and Zombie Yellow™ emits into the Brilliant Violet 570™ channel.

Analysis of Dead and Apoptotic splenocytes with Zombie Aqua™ and Annexin V.

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Zombie Aqua™/ Zombie NIR™ /Zombie Yellow™

Q: Why can’t I fix my cells prior to using Zombie Aqua™/ Zombie NIR™/Zombie Yellow™?A: The fixation process can contort and alter the membrane of cells, effectively rendering them as dead. Since the ability of these dyes to stain dead cells is correlated with cell permeability, your results may no longer be a valid representation of dead versus live cells.Q: Can I use methanol/ethanol for fixation after using Zombie Aqua™/Zombie NIR™/Zombie Yellow™?A: Yes, most fixation reagents are fine to be used with Zombie Aqua™, Zombie NIR™ and Zombie Yellow™. However, it should be noted that they can still be sensitive to reactive oxygen species. Light exposure or reagents with hydrogen peroxide can lead to free radical formation, affecting fluorescence.Q: How does the performance of your Zombie Aqua™/ Zombie NIR™/Zombie Yellow™ compare with competitors?A: Zombie Aqua™, Zombie NIR™ and Zombie Yellow™ have been tested against other leading competitors’ fixable viability kits and given comparable results. We also highly recommend that you titrate down the amount of each dye used in order to best match the negative signals of your unstained sample and MFI- (mean fluorescence intensity) stained samples.

Q: Can I use the UV laser to stimulate Zombie Aqua™? If so, can I then use it in conjunction with BV510™?A: While we typically do not test Zombie Aqua™ with the UV laser, its excitation peak suggests it is effectively excited at 355 nm. However, we would not recommend using BV510™ off the violet laser and Zombie Aqua™ off the UV laser at the same time. Due to cross-beam excitation of BV510™ by the UV laser and the violet excitation of Zombie Aqua™, this would lead to significantly increased background and excessive compensation requirements.Q: Can I use Zombie Aqua™/Zombie NIR™/Zombie Yellow™ and Annexin V to detect apoptotic cells?A: Yes, they can be used with Annexin V to discriminate live, apoptotic, and dead cells. Cells double positive for both Zombie and Annexin V are dead, while Zombie-dim/Annexin V-positive cells are apoptotic. Live cells will be Zombie-low and Annexin V-negative. The advantage to Zombie dyes over PI and 7-AAD is that you can now fix and/or permeabilize the cells to stain for cell surface and intracellular antigens.

FAQs

NameExcitation (max)

Emission (max) Size Cat. No.

Zombie Aqua™ Fixable Viability Kit 382 nm 510 nm 100 tests | 500 tests 423101 | 423102

Zombie NIR™ Fixable Viability Kit 718 nm 746 nm 100 tests | 500 tests 423105 | 423106

Zombie Yellow™ Fixable Viability Kit 396 nm 572 nm 100 tests | 500 tests 423103 | 423104

Propidium Iodide Solution 540 nm 620 nm 2 mL (200 tests) 421301

7-AAD Viability Staining Solution 550 nm 645 nm 200 tests | 500 tests 420403 | 420404

Product List

Plan

* I plan to analyze my cells without fixation or sort my cells.

* I plan to fix my cells prior to analysis.

Options

Propidium Iodide, 7-AAD, or Zombie Aqua™/Zombie NIR™/Zombie Yellow™

Zombie Aqua™/ Zombie NIR™/Zombie Yellow™

Strategy Guide

Note: We recommend the Zombie Aqua™/ Zombie NIR™ /Zombie Yellow™ be used first, prior to addition of antibodies or fixation of cells.

302

Buffers, Solutions, and Chemicals

Toll Free 1.877.BIOLEGEND (246.5343)

Description Applications Cat. No. Size Price ($)

7-AAD Viability Staining Solution FC 420403 | 420404 200 tests | 500 tests 25 | 50

Accutase® Cell Detachment Solution FC 423201 100 mL 29

Annexin V Binding Buffer FC, IF 422201 100 mL 30

Brefeldin A Solution (1,000X) ICFC 420601 1 mL 55

Cell Staining Buffer FC, ICC, ICFC 420201 500 mL 75

CFSE ICFC, IF, in vivo cell tracking 422701 500 μg 85

Cyto-Last™ Buffer ICFC 422501 100 mL 65

DAPI (4',6-Diamidino-2-Phenylindole, Dilactate) ICFC, IF 422801 10 mg 85

ELISA Assay Diluent (5X) ELISA 421203 100 mL 60

ELISA Coating Buffer (5X) ELISA 421701 30 mL 15

ELISA Wash Buffer (20X) ELISA 421601 500 mL 120

Fixation Buffer ICFC, ICC 420801 100 mL 50

FluoroFix™ Buffer FC, ICC, ICFC 422101 200 tests 130

FOXP3 Fix/Perm Buffer (4X) ICFC 421401 30 mL 60

FOXP3 Fix/Perm Buffer Set ICFC 421403 100 tests 120

FOXP3 Perm Buffer (10X) ICFC 421402 25 mL 60

Human TruStain FcX™ (Fc Receptor Blocking Solution) FC, ICFC, ICC 422301 | 422302 50 tests | 200 tests 85 | 195

Monensin Solution (1,000X) ICFC 420701 1 mL 45

Nuclear Factor Fixation and Permeabilization Buffer Set ICFC 422601 100 tests 120

Permeabilization Wash Buffer (10X) ICC, ICFC, IHC 421002 100 mL 80

Propidium Iodide Solution FC 421301 2 mL 45

Rainbow Calibration Particles, 6 peaks (3.0-3.4 μm) FC 422901 5 mL 255

Rainbow Calibration Particles, 8 peaks (3.0-3.4 μm) FC 422903 5 mL 250

Rainbow Fluorescent Particles, 1 peak (3.0-3.4 μm) - Bright Intensity FC 422907 5 mL 245

Rainbow Fluorescent Particles, 1 peak (3.0-3.4 μm) - Mid Range Intensity FC 422905 5 mL 245

RBC Lysis Buffer (10X) FC, ICFC 420301 100 mL 55

RBC Lysis/Fixation Solution (10X) FC 422401 50 mL 120

Stop Solution for TMB Substrate ELISA 423001 60 mL 30

TMB High Sensitivity Substrate Solution ELISA 421501 60 mL 25

TMB Substrate Reagent Set ELISA 421101 100 mL 35

Zombie Aqua™ Fixable Viability Kit FC, ICFC 423101 | 423102 100 tests | 500 tests 75 | 285

Zombie NIR™ Fixable Viability Kit FC, ICFC 423105 | 423106 100 tests | 500 tests 75 | 285

Zombie Yellow™ Fixable Viability Kit FC, ICFC 423103 | 423104 100 tests | 500 tests 75 | 285