FITC Annexin V

FITC Annexin V
Catalog# / Size 640905 / 25 tests
640906 / 100 tests
640945 / 300 tests
Other Names Annexin A5
Description 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 cells
and necrotic. 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
Reactivity All mammalian species
Formulation Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 0.2% (w/v) BSA (origin
USA).
Preparation The purified protein was conjugated with FITC under optimal conditions.
Concentration Lot-specific (please contact technical support for concentration and total µg amount, or use our Lookup
tool if you have a lot number.)
Storage & Handling The Annexin V solution should be stored undiluted between 2°C and 8°C, and protected from
prolonged exposure to light. Do not freeze.
Application FC - Quality tested
Recommended Usage Each lot of this product is quality control tested by immunofluorescent staining with flow cytometric
analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl 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.
Excitation Laser Blue Laser (488 nm)
Application Notes Annexin V Staining
1. Wash cells twice with cold BioLegend cell staining buffer (cat # 420201) and then resuspend cells in
Annexin V Binding Buffer (cat # 422201) at a concentration of 1x10e6 cells/ml.
2. Transfer 100 µl of cell suspension in 5 ml test tube.
3. Add 5 µl of FITC Annexin V.
4. Add 10 µl of PI solution (cat # 421301) or 7-AAD (cat # 420403/420404).
5. Gently vortex the cells and incubate for 15 min at RT (25 °C) in the dark.
6. Add 400 µl of Annexin V Binding Buffer (cat # 422201) to each tube. Analyze by flow cytometry.
Application References
(PubMed link indicates
BioLegend citation)
1. Koopman G, et al. 1994. Blood 84:1415.
2. Vermes I, et al. 1995. J. Immunol. Methods 184:39.
3. Dachary-Prigent J, et al. 1993. Blood 81:2554.
4. Sekine C, et al. 2009. Int Immunol. PubMed
5. Grujic M, et al. 2010. J. Immunol. 185:1730. PubMed
6. Mulik RS, et al. 2010. Int J Pharm. 398:190. PubMed
7. Gupta A, et al. 2011. Leuk Res. 35:1498. PubMed
8. Speth C,et al. 2013. J Infect Dis. 207:823. PubMed
9. Wang Q, et al. 2013. Genes Dev. 27:615. PubMed
10. Gill K, et al. 2013. Biocheim Biophys Acta. 830:2763. PubMed
11. Juel HB, et al. 2013. PLoS One. 8:64619. PubMed
12. Hellmann J, et al. 2013. J. Immunol. 191:1383. PubMed
Product Citations
1. Sekine C, et al. 2009. Int Immunol. 1.079861111. PubMed
2. Grujic M, et al. 2010. J Immunol. 185:1730. PubMed
3. Putheti P, et al. 2010. PLoS One. 5:e8706. PubMed
4. Speth C, et al. 2013. J Infect Dis. 207:823. PubMed
5. Wang Q, et al. 2013. Genes Dev. 27:615. PubMed
6. Gill K, et al. 2013. Biochim Biophys Acta. 1830:2763. PubMed
7. Juel H, et al. 2013. PLoS One. 8:64619. PubMed
Version: 4 Revision Date: 07/01/2016 
8. Gupta A, et al. 2011. Leuk Res. 35:1498. PubMed
9. Mulik R, et al. 2010. Int J Pharm. 398:190. PubMed
10. Hellmann J, et al. 2013. J Immunol. 191:1383. PubMed
11. Séguier S, et al. 2013. PLoS One. 8:70937. PubMed
12. Passaro C, et al. 2013. Endocr Relat Cancer. 20:633. PubMed
RRID not an antibody (BioLegend Cat. No. 640905)
not an antibody (BioLegend Cat. No. 640906)
not an antibody (BioLegend Cat. No. 640945)
Antigen Details
Biology Area Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Neuroscience
Gene ID 308
Product Data
Human T leukemia cell line, Jurkat, nontreated
(top) or treated (bottom) with
BioLegend’s anti-human CD95 (EOS9.1)
mAb (cat. 305704) for 6 hours, then
stained with Annexin V-FITC and
Propidium Iodide (PI) (cat. 421301)
For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other
violations that may occur with the use of our products.
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website,
www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to
manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written
approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses.
Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or
commercial use.
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