Annexin V Binding Buffer

Pricing & Availability
Other Names
Annexin-V-Binding Buffer
Ave. Rating
Submit a Review
Product Citations
Cat # Size Price Quantity Avail. Save
422201 100 mL $30
Check Availability

Need larger quantities of this item?
Request Bulk Quote

Annexin V Binding Buffer has been formulated for flow cytometric labeling of apoptotic cells with Annexin V reagents.  The buffer is provided as a ready-to-use solution.

Product Details
Technical data sheet

Product Details

Storage & Handling
Store between 2°C and 8°C.

FC - Quality tested
IF - Reported in the literature, not verified in house

Recommended Usage

Annexin V Binding Buffer is recommended for use with Annexin V reagents. Cells should be resuspended in Annexin V Binding Buffer at a concentration 1x106/ml and then combined with Annexin V reagents at the recommended volumes.

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 1x106 cells/ml.
2. Transfer 100 µl of cell suspension in 5 ml test tube.
3. Add 5 µl of fluorochrome conjugated 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.

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)
  1. Vermes I, et al. 1995. J. Immunol. Methods 184:39.
  2. Soon PS, et al. 2013. Endocr Relat Cancer. 20:1. PubMed
  3. Turner JE, et al. 2013. J Exp Med. PubMed
  4. Lindner JM, et al. 2013. Mol Cell Biol. 33:4628. PubMed
  5. Turner JE, et al. 2013 J. Exp Med. 210:2951. PubMed
  6. Beggs KM, et al. 2014. Toxicol Sci. 137:91. PubMed
  7. Miyazawa M, et al. 2014. Mol Biol Cell. 25:2116. PubMed
  8. Burbulla LF, et al. 2014. Cell Death Dis. 5:1180. PubMed
  9. de Vires M, 2014. Am J Physiol Lung Cell Mol Physiol. 307:240. PubMed
  10. Sahin E, 2014. J. Immunol. 193:1717. PubMed
  11. Xiong R, et al. 2014. Toxicol Appl Pharmacol. 280:285. PubMed
  12. Thakran S, 2015. Invest Ophthalmol Vis Sci. 56:177. PubMed
Product Citations
  1. Soon P, et al. 2013. Endocr Relat Cancer. 20:1. PubMed
  2. Lindner J, et al. 2013. Mol Cell Biol. 33:4628. PubMed
  3. Turner J, et al. 2013. J Exp Med. 210:2951. PubMed
  4. Beggs K, et al. 2014. Toxicol Sci. 137:91. PubMed
  5. Tsuji M 2014. Mol Biol Cell. 25:2116. PubMed
  6. Burbulla L, et al. 2014. Cell Death Dis. 5:1180. PubMed
  7. Vries M, et al. 2014. Am J Physiol Lung Cell Mol Physiol. 307:240. PubMed
  8. Sahin E, et al. 2014. J Immunol. 193:1717. PubMed
  9. Xiong R, et al. 2014. Toxicol Appl Pharmacol. 280:285. PubMed
  10. Thakran S, et al. 2015. Invest Ophthalmol Vis Sci . 56:177. PubMed
  11. Kojima N, et al. 2015. Anticancer Res. 35: 4493-4499. PubMed
  12. Cabet E, et al. 2015. PLoS One. 10: 0137009. PubMed
  13. Xiong R, et al. 2015. Mol Pharmacol. 88: 1045 - 1054. PubMed
  14. Suarez S, et al. 2015. J Biol Chem. 290: 28311 - 28320. PubMed
  15. Hopkins T, et al. 2015. Nucleic Acids Res. 10.1093/nar/gkv1515. PubMed
  16. Yabas M, et al. 2016. PLoS One. 11: 0146774. PubMed
  17. Deventhiran J, et al. 2016. J Virol. 90: 222 - 231. PubMed
  18. Itkin T, et al. 2016. Nature. 532: 323-328. PubMed
  19. Hickman O, et al. 2016. Br J Cancer. 10.1038/bjc.2016.91. PubMed
  20. Peiris T, et al. 2016. Development. 143: 1697 - 1709.. PubMed
  21. Hergott C, et al. 2016. Blood. 127: 2460 - 2471. PubMed
  22. Soprano M, et al. 2016. PLoS One. 11: 0155970. PubMed
  23. Langsch S, et al. 2016. Cancer Res . 76: 4160 - 4169. PubMed
  24. Li P, et al. 2016. Mol Biol Cell. 27: 2542 - 2553. PubMed
  25. Becker A, et al. 2016. Sci Rep. 6: 33847. PubMed
  26. Li L, et al. 2016. Cell Death Dis. 7:e2439. PubMed
  27. L Chang, Y Li, D Kaplan 2016. Sci Rep. 6:36862. PubMed
  28. Guo H, et al. 2016. Sci Rep. 6:37559. PubMed
  29. Guentsch A, et al. 2017. Mol Cell Biol. 37: e00236-16. PubMed
  30. Kitazawa M, et al. 2017. PLoS One. 10.1371/journal.pone.0169340. PubMed
  31. Herold N, et al. 2017. Nat Med. 23:256-263. PubMed
  32. Domenis R, et al. 2017. PLoS One. 10.1371/journal.pone.0169932. PubMed
  33. Marton N, et al. 2017. Cellular and Molecular Life Sciences. 10.1007/s00018-017-2535-8. PubMed
  34. Terashima S, et al. 2017. J Radiat Res. 10.1093/jrr/rrx025. PubMed
  35. Krishnasamy K,et al. 2017. Nat Commun. . 10.1038/s41467-017-00953-2. PubMed
  36. Shrestha C,et al. 2017. PLoS One. 10.1371/journal.pone.0186169. PubMed
  37. Naaman H,et al. 2017. PLoS One. 10.1371/journal.pone.0187077. PubMed
  38. Hönes J, et al. 2017. Sci Rep. . 10.1038/s41598-017-15866-9. PubMed
  39. Feng S, et al. 2017. Cancer Sci. 108:12:00. PubMed
  40. Litzenburger UM, et al. 2017. Genome Biol. 18:15. PubMed
  41. Zhou L, et al. 2017. Tumour Biol. 39:1010428317694313. PubMed
  42. Jung E, et al. 2017. J Neurosci. 37:6837. PubMed
  43. Kranz P, et al. 2017. Cell Death Dis. 8:e2986. PubMed
  44. Venter C, et al. 2017. Cell Physiol Biochem. 43:1288. PubMed
  45. Lee HW, et al. 2018. Am J Physiol Renal Physiol. 314:F280. PubMed
  46. Lim JA, et al. 2018. BMC Cancer. 0.860416667. PubMed
  47. Saint Fleur–Lominy S, et al. 2018. Cell Rep. 24:3045. PubMed
  48. Li B, et al. 2019. Oncol Rep. 41:608. PubMed
  49. Wang M, et al. 2018. Diabetologia. 62:335. PubMed
  50. Zhang S, et al. 2018. Sci Rep. 8:17066. PubMed
  51. Kim J, et al. 2019. PLoS One. 14:e0210248. PubMed
  52. Rosenberger L, et al. 2019. Sci Rep. 0.835416667. PubMed
  53. Wang P, et al. 2019. Nat Commun. 0.940972222. PubMed
  54. Pedro L, et al. 2019. Small. 15:e1900224. PubMed
  55. Sun T, et al. 2019. Cancer Med. 2.457638889. PubMed
  56. Hasselluhn MC, et al. 2019. Cell Death Dis. 0.726388889. PubMed
  57. Vancsik T, et al. 2019. Cancer Med. 3.313888889. PubMed
  58. Djeungoue–Petga MA, et al. 2019. Cell Death Dis. 1.069444444. PubMed
  59. Bersini S, et al. 2020. Life Sci Alliance. 3:e201900623. PubMed
  60. Moose DL, et al. 2020. Cell Rep. 30:3864. PubMed
  61. Quan M, et al. 2019. Oncol Rep. 41:3051. PubMed
  62. Luca E, et al. 2020. Mol Metab. 36:100967. PubMed

Antigen Details

Gene ID

Related FAQs

Can I freeze Annexin V conjugates?

It should not be frozen as it will lead to loss of biological activity due to dimerization.

Can I use RPMI during Annexin V staining?

It is best to follow protocol as described on the product data sheet. Moreover, RPMI 1640 has a relatively high concentration of phosphate and low calcium ion concentration, which negatively impacts Annexin binding to its target phosphatidylserine (PS). Measurement of cell death by using Annexin V may also be significantly affected by time of incubation on ice, calcium concentration, and type of medium.

How does pH and staining temperature affect Annexin V-Phosphatidylserine binding?

Annexin-Phosphatidylserine binding is lost below pH 5.2 and with prolonged incubation over a temperature of 42°C.

How is your Annexin made and what sequence does it cover?

It is made in E. coli, covering human aa Met1-Asp320.

Is Annexin V suitable for conjugation with the Maxpar® kit for CyTOF®?

Maxpar® Labeling kits require the protein to be partially reduced, so the metal chelate can be introduced through an SH group in the hinge region of the reduced antibody. Human Annexin V contains only one Cysteine which was reported to be chemically inactive. Thus, the Maxpar® labeling protocol would not work with Annexin V, unless a free –SH group can be introduced to Annexin V.  For more information regarding SH-mediated conjugation of Annexin V please consult published papers such as this one.

Why do I need to use Annexin V Binding Buffer?

Annexin V binding requires the presence of calcium in the solution.  So, we provide Annexin V Binding Buffer (cat # 422201), which is optimized for the best performance of Annexin V staining.

Go To Top Version: 4    Revision Date: 05/20/2020

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, 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.


8999 BioLegend Way, San Diego, CA 92121
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

Insert Note Here
Save Close Clear
Lab Timer
Forgot your password? Reset Password
Request an Account