- Regulatory Status
- RUO
- Other Names
- Annexin A5 Apoptosis Detection Kit
Cat # | Size | Price | Quantity Check Availability | ||
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640932 | 100 tests | $267.00 |
BioLegend's APC Annexin V Apoptosis Detection Kit with PI has been specifically designed for the identification of apoptotic and necrotic cells.
Annexin V (or Annexin A5) is a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner. PS is normally only found on the intracellular leaflet of the plasma membrane in healthy cells, but during early apoptosis, membrane asymmetry is lost and PS translocates to the external leaflet. Fluorochrome-labeled Annexin V can then be used to specifically target and identify apoptotic cells. Annexin V Binding Buffer is recommended for use with Annexin V staining. Annexin V binding alone cannot differentiate between apoptotic and necrotic cells. To help distinguish between the necrotic and apoptotic cells we recommend use of our Propidium Iodide Solution (PI). Early apoptotic cells will exclude PI, while late stage apoptotic cells and necrotic cells will stain positively, due to the passage of these dyes into the nucleus where they bind to DNA.
Propidium iodide is a fluorescent dye that binds to DNA. When excited by 488 nm laser light, it can be detected with in the PE/Texas Red® channel with a bandpass filter 610/10. It is commonly used in evaluation of cell viability or DNA content in cell cycle analysis by flow cytometry.
Product Details
- Verified Reactivity
- Human, Mouse, Rat
- Reported Reactivity
- Other Species
- Concentration
- Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
- Storage & Handling
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Store between 2°C and 8°C. Do not freeze.
Caution: Propidium Iodide Solution is toxigenic and mutagenic; handle with care. - Application
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FC - Quality tested
- Recommended Usage
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Staining Procedure:
1. Wash cells twice with cold BioLegend's Cell Staining Buffer, and then resuspend cells in Annexin V Binding Buffer at a concentration of 0.25-1.0 x 107 cells/mL.
2. Transfer 100 µL of cell suspension in a 5 mL test tube.
3. Add 5 µL of APC Annexin V.
4. Add 10 µL of Propidium Iodide Solution.
5. Gently vortex the cells and incubate for 15 min at room temperature (25°C) in the dark.
6. Add 400 µL of Annexin V Binding Buffer to each tube. Analyze by flow cytometry with proper machine settings. - Application Notes
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Materials Provided:
0.5 ml of APC Annexin V
1 ml of Propidium Iodide Solution
50 ml of Annexin V Binding Buffer
Materials Not Included:
Cell Staining Buffer (Cat. No. 420201)
For a better experience detecting apoptosis, we now recommend Apotracker™. Cell staining with Apotracker™ is Calcium independent. Thus, no special buffers are required, and the protocol can be shortened for single-step co-staining with other reagents. -
Application References
(PubMed link indicates BioLegend citation) - Product Citations
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Antigen Details
- Biology Area
- Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Proliferation and Viability, Neuroscience
- Gene ID
- 308 View all products for this Gene ID