Annexin V Binding Buffer

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Regulatory Status
RUO
Other Names
Annexin-V-Binding Buffer
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422201 100 mL $30.00
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Description

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.
Application

FC - Quality tested

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. Marton N, et al. 2017. Cellular and Molecular Life Sciences. 10.1007/s00018-017-2535-8. PubMed
  2. Terashima S, et al. 2017. J Radiat Res. 10.1093/jrr/rrx025. PubMed
  3. Krishnasamy K,et al. 2017. Nat Commun. . 10.1038/s41467-017-00953-2. PubMed
  4. Shrestha C,et al. 2017. PLoS One. 10.1371/journal.pone.0186169. PubMed
  5. Naaman H,et al. 2017. PLoS One. 10.1371/journal.pone.0187077. PubMed
  6. Hönes J, et al. 2017. Sci Rep. . 10.1038/s41598-017-15866-9. PubMed
  7. Feng S, et al. 2017. Cancer Sci. 108:12:00. PubMed
  8. Litzenburger UM, et al. 2017. Genome Biol. 18:15. PubMed
  9. Zhou L, et al. 2017. Tumour Biol. 39:1010428317694313. PubMed
  10. Jung E, et al. 2017. J Neurosci. 37:6837. PubMed
  11. Kranz P, et al. 2017. Cell Death Dis. 8:e2986. PubMed
  12. Venter C, et al. 2017. Cell Physiol Biochem. 43:1288. PubMed
  13. Lee HW, et al. 2018. Am J Physiol Renal Physiol. 314:F280. PubMed
  14. Lim JA, et al. 2018. BMC Cancer. 0.860416667. PubMed
  15. Saint Fleur–Lominy S, et al. 2018. Cell Rep. 24:3045. PubMed
  16. Li B, et al. 2019. Oncol Rep. 41:608. PubMed
  17. Wang M, et al. 2018. Diabetologia. 62:335. PubMed
  18. Zhang S, et al. 2018. Sci Rep. 8:17066. PubMed
  19. Kim J, et al. 2019. PLoS One. 14:e0210248. PubMed
  20. Rosenberger L, et al. 2019. Sci Rep. 0.835416667. PubMed
  21. Wang P, et al. 2019. Nat Commun. 0.940972222. PubMed
  22. Pedro L, et al. 2019. Small. 15:e1900224. PubMed
  23. Sun T, et al. 2019. Cancer Med. 2.457638889. PubMed
  24. Hasselluhn MC, et al. 2019. Cell Death Dis. 0.726388889. PubMed
  25. Vancsik T, et al. 2019. Cancer Med. 3.313888889. PubMed
  26. Djeungoue–Petga MA, et al. 2019. Cell Death Dis. 1.069444444. PubMed
  27. Bersini S, et al. 2020. Life Sci Alliance. 3:e201900623. PubMed
  28. Moose DL, et al. 2020. Cell Rep. 30:3864. PubMed
  29. Quan M, et al. 2019. Oncol Rep. 41:3051. PubMed
  30. Luca E, et al. 2020. Mol Metab. 36:100967. PubMed
  31. Labi V, et al. 2019. Genes Dev. 33:1673. PubMed
  32. Soon P, et al. 2013. Endocr Relat Cancer. 20:1. PubMed
  33. Lindner J, et al. 2013. Mol Cell Biol. 33:4628. PubMed
  34. Turner J, et al. 2013. J Exp Med. 210:2951. PubMed
  35. Beggs K, et al. 2014. Toxicol Sci. 137:91. PubMed
  36. Tsuji M 2014. Mol Biol Cell. 25:2116. PubMed
  37. Burbulla L, et al. 2014. Cell Death Dis. 5:1180. PubMed
  38. Vries M, et al. 2014. Am J Physiol Lung Cell Mol Physiol. 307:240. PubMed
  39. Sahin E, et al. 2014. J Immunol. 193:1717. PubMed
  40. Xiong R, et al. 2014. Toxicol Appl Pharmacol. 280:285. PubMed
  41. Thakran S, et al. 2015. Invest Ophthalmol Vis Sci . 56:177. PubMed
  42. Kojima N, et al. 2015. Anticancer Res. 35: 4493-4499. PubMed
  43. Cabet E, et al. 2015. PLoS One. 10: 0137009. PubMed
  44. Xiong R, et al. 2015. Mol Pharmacol. 88: 1045 - 1054. PubMed
  45. Suarez S, et al. 2015. J Biol Chem. 290: 28311 - 28320. PubMed
  46. Hopkins T, et al. 2015. Nucleic Acids Res. 10.1093/nar/gkv1515. PubMed
  47. Yabas M, et al. 2016. PLoS One. 11: 0146774. PubMed
  48. Deventhiran J, et al. 2016. J Virol. 90: 222 - 231. PubMed
  49. Itkin T, et al. 2016. Nature. 532: 323-328. PubMed
  50. Hickman O, et al. 2016. Br J Cancer. 10.1038/bjc.2016.91. PubMed
  51. Peiris T, et al. 2016. Development. 143: 1697 - 1709.. PubMed
  52. Hergott C, et al. 2016. Blood. 127: 2460 - 2471. PubMed
  53. Soprano M, et al. 2016. PLoS One. 11: 0155970. PubMed
  54. Langsch S, et al. 2016. Cancer Res . 76: 4160 - 4169. PubMed
  55. Li P, et al. 2016. Mol Biol Cell. 27: 2542 - 2553. PubMed
  56. Becker A, et al. 2016. Sci Rep. 6: 33847. PubMed
  57. Li L, et al. 2016. Cell Death Dis. 7:e2439. PubMed
  58. L Chang, Y Li, D Kaplan 2016. Sci Rep. 6:36862. PubMed
  59. Guo H, et al. 2016. Sci Rep. 6:37559. PubMed
  60. Guentsch A, et al. 2017. Mol Cell Biol. 37: e00236-16. PubMed
  61. Kitazawa M, et al. 2017. PLoS One. 10.1371/journal.pone.0169340. PubMed
  62. Herold N, et al. 2017. Nat Med. 23:256-263. PubMed
  63. Domenis R, et al. 2017. PLoS One. 10.1371/journal.pone.0169932. PubMed
  64. Leiendecker L, et al. 2020. EMBO Mol Med. 12:e12525. PubMed
  65. Liu M, et al. 2021. Mol Med Rep. 23:00. PubMed
  66. Zhang S, et al. 2020. Front Immunol. 1.757638889. PubMed
  67. Cheng CC, et al. 2020. Elife. 9:00. PubMed
  68. Verma N, et al. 2020. Sci Adv. 6:00. PubMed
  69. Festag J, et al. 2020. Mol Ther Nucleic Acids. 1.330555556. PubMed
  70. Hagan AS, et al. 2020. Development. 147:00:00. PubMed
  71. Xie S, et al. 2021. Sci Rep. 0.660416667. PubMed

Antigen Details

Gene ID
NA

Related FAQs

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

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

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.

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.

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.

Can I freeze Annexin V conjugates?

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

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.

Go To Top Version: 5    Revision Date: 03/15/2021

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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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.
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