- Regulatory Status
- RUO
- Other Names
- Annexin A5
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Human T leukemia cell line Jurkat, treated (left) or non-treated (right) with BioLegend's anti-human CD95 (EOS9.1) mAb (cat. 305704) for 4 hours at 37°C, then stained with Annexin V- APC/Fire™ 750 and Helix NP Green (cat. 425303 at 1.25nM) for 15 minutes at 37°C in Annexin Binding buffer.
| Cat # | Size | Price | Quantity Check Availability | ||
|---|---|---|---|---|---|
| 640952 | 25 tests | $136.00 | |||
| 640953 | 100 tests | $347.00 | |||
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 (Cat. No. 422201) is recommended for use with Annexin V staining. Annexin V binding alone cannot differentiate between apoptotic and necrotic cells. Therefore, we recommend using our Helix NP™ Blue (Cat. No. 425305), Helix NP™ Green (Cat. No. 425303) or Helix NP™ NIR (Cat. No. 425301). Early apoptotic cells will exclude 7-AAD and 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.
Product Details
- Verified Reactivity
- Human, Mouse, Rat
- Reported Reactivity
- Other Species
- Formulation
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
- Preparation
- The antibody was purified by affinity chromatography and conjugated with APC/Fire™ 750 under optimal conditions.
- Concentration
- Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
- Storage & Handling
- The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
- Application
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FC - Quality tested
- Recommended Usage
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Each lot of this product is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric application, the suggested use of this reagent is 5 - 15 µg per 100,000 - million cells in a 100 µL volume of Annexin V Binding Buffer (Cat No. 422201). It is recommended that the reagent be titrated for optimal performance for each application.
* APC/Fire™ 750 has a maximum excitation of 650 nm and a maximum emission of 774 nm. - Excitation Laser
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Red Laser (633 nm)
- Application Notes
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Annexin V Staining
- Wash cells twice with cold BioLegend Cell Staining Buffer (Cat. No. 420201) and then resuspend cells in Annexin V Binding Buffer (Cat. No. 422201) at a concentration of 1x106 cells/mL.
- Transfer 100 µL of cell suspension in 5 mL test tube.
- Add 5 µL of fluorochrome conjugated Annexin V.
- Stain with a viability dye, such as PI (Cat. No. 421301), 7-AAD (Cat. Nos. 420403 & 420404), or Helix NP dyes (Cat. Nos. 425301, 425303, & 425305), if desired.
- Gently vortex the cells and incubate for 15 min at RT (25°C) in the dark.
- Add 400* µL of Annexin V Binding Buffer (Cat. No. 422201) to each tube. *For more concentrated samples, add a minimum of 200 µl of Annexin V Binding Buffer in this step.
- Analyze by flow cytometry.
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.
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Application References
(PubMed link indicates BioLegend citation) - Product Citations
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- RRID
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not an antibody (BioLegend Cat. No. 640952)
not an antibody (BioLegend Cat. No. 640953)
Antigen Details
- Biology Area
- Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Neuroscience
- Gene ID
- 308 View all products for this Gene ID
- UniProt
- View information about Annexin V on UniProt.org