Metabolic Activity Dead Cell Apoptosis Kit with C12 Resazurin, Annexin V APC, and SYTOX™ Green, for flow cytometry
Metabolic Activity Dead Cell Apoptosis Kit with C12 Resazurin, Annexin V APC, and SYTOX™ Green, for flow cytometry
Invitrogen™

Metabolic Activity Dead Cell Apoptosis Kit with C12 Resazurin, Annexin V APC, and SYTOX™ Green, for flow cytometry

This flow cytometry product provides a three-color fluorescence assay that distinguishes live, apoptotic, and late apoptotic cells from one another.Read more
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Catalog number V35114
Price (USD)
964.00
Each
Add to cart
Price (USD)
964.00
Each
Add to cart
This flow cytometry product provides a three-color fluorescence assay that distinguishes live, apoptotic, and late apoptotic cells from one another. These populations can easily be distinguished using a flow cytometer that has both 488 nm and 633 nm excitation sources (an argon-ion laser and a HeNe laser) and the following reagents Annexin V to detect phosphatidylserine, C12 resazurin for cell metabolism, and SYTOX™ Green nucleic acid stain for compromised membranes.

View a selection guide for all apoptosis assays for flow cytometry.
WARNING: Cancer and Reproductive Harm – www.P65Warnings.ca.gov
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Quantity1 kit
FormatTube, Slide
Product TypeDead Cell Apoptosis Kit
Flow Cytometer Laser Lines633/635, 488
Excitation/EmissionC12-Resazurin: 571⁄585, SYTOX™ green: 503⁄524, APC: 650⁄660
No. of Reactions50 reactions
Shipping ConditionWet Ice
Product LineSYTOX™
For Use With (Application)Flow Cytometry
ConjugateAPC, SYTOX Green, C12-resazurin
For Use With (Equipment)Fluorescence Microscope, Flow Cytometer
Unit SizeEach
Contents & Storage
Contains 1 vial of annexin V, APC conjugate (250 μL), 1 vial of SYTOX™ green stain (100 μL), 1 vial of C12-resazurin (40 μg), and 1 vial of DMSO (1.5 mL).

Store in refrigerator (2–8°C) and protect from light.

Frequently asked questions (FAQs)

I trypsinized my adherent cells and labeled with annexin V, and now my flow data is showing a high percentage of apoptotic cells even for control, untreated cells. What is the problem?

Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. Allow the cells to recover for about 30 minutes in optimal cell culture conditions and medium after trypsinizing/scraping so that they can recover their membrane integrity before staining. For lightly adherent cell lines, such as HeLa and NIH 3T3, another option is to use non-enzyme treatments like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I detect annexin V staining in an imaging assay?

Annexin V staining is not typically used in imaging experiments; it is a better reagent for flow cytometry analysis. All cells will stain to some extent, so it can be difficult to distinguish a relatively bright annexin V-stained cell from a dimmer non-apoptotic cell. Caspase activation, detected using our CellEvent Caspase 3/7 or Image-iT LIVE Caspase detection kits, is a better method for detecting apoptosis in an imaging assay.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

When should I stain adherent cells with annexin V for flow cytometric analysis? Before or after I trypsinize them?

Trypsinize first and then allow the cells to recover about 30 minutes in optimal cell culture conditions and medium before staining with annexin V conjugates. Trypsinization or mechanical scraping of cells temporarily disrupts the plasma membrane, allowing for annexin V to bind phosphatidylserine on the cytoplasmic surface of the cell membrane and thus leading to false positive staining. For lightly adherent cell lines such as HeLa and NIH 3T3, you could use a less harsh (non-enzymatic) dissociation product like Gibco Cell Dissociation Buffer (Cat. No. 13151014).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can I fix my cells after annexin V labeling?

Annexin V staining is best analyzed on live cells. If you need to fix your cells for analysis, then fix in 3.7% formaldehyde in PBS containing calcium and magnesium to maintain binding during fixation. The signal will not be retained after permeabilization, thus annexin V staining is not compatible with internal antibody labeling.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

How do alamarBlue reagent and PrestoBlue reagent differ from resazurin and C12-resazurin?

alamarBlue reagent and PrestoBlue reagent contain resazurin in a proprietary stabilizing formulation that allows for a convenient “mix, incubate, and read” protocol. PrestoBlue reagent is an improvement in the formulation of alamarBlue reagent that allows for much faster staining (typically 10 minutes vs. 1-4 hours to obtain a similar signal and sensitivity). C12-resazurin is a derivative of resazurin that has better cellular retention and thus allows for analysis on a flow cytometer and multiplexing with viability indicators and other biomarkers.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.