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Annexin V Binding Buffer

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Regulatory Status
RUO
Other Names
Annexin-V-Binding Buffer
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422201 100 mL $36
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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. No. 420201) and then resuspend cells in Annexin V Binding Buffer (Cat. No. 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. 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. 
  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. No. 422201) to each tube. *For more concentrated samples, add a minimum of 200 µl of Annexin V Binding Buffer in this step.
  7. 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. Tsuji M 2014. Mol Biol Cell. 25:2116. PubMed
  2. Guo H, et al. 2016. Sci Rep. 6:37559. PubMed
  3. Kitazawa M, et al. 2017. PLoS One. 10.1371/journal.pone.0169340. PubMed
  4. Liu M, et al. 2021. Mol Med Rep. 23:00. PubMed
  5. Ocklenburg T, et al. 2021. Sci Rep. 7199:11. PubMed
  6. Shrestha C,et al. 2017. PLoS One. 10.1371/journal.pone.0186169. PubMed
  7. Djeungoue–Petga MA, et al. 2019. Cell Death Dis. 1.069444444. PubMed
  8. Forika G, et al. 2021. Pathol Oncol Res. 27:1610048. PubMed
  9. Chen Q, et al. 2022. Cell Rep. 39:110990. PubMed
  10. Clusmann J, et al. 2022. Cell Death Dis. 13:702. PubMed
  11. Hao J, et al. 2022. Cell Rep. 41:111804. PubMed
  12. Shahid AM, et al. 2022. PLoS One. 17:e0278209. PubMed
  13. Cappellesso F, et al. 2022. Nat Cancer. 3:1464. PubMed
  14. Muckenhuber M, et al. 2023. Am J Transplant. 23:84. PubMed
  15. Zhao Q, et al. 2023. Nat Commun. 14:735. PubMed
  16. Wang YH, et al. 2022. Sci Adv. 8:eabn6552. PubMed
  17. Vajda F, et al. 2022. Cells. 11:. PubMed
  18. Chu CY, et al. 2023. Sci Rep. 13:3943. PubMed
  19. Riera-Domingo C, et al. 2023. Sci Adv. 9:eadd5028. PubMed
  20. Bujarrabal-Dueso A, et al. 2023. Nat Struct Mol Biol. 30:475. PubMed
  21. Quintela M, et al. 2023. Clin Epigenetics. 15:63. PubMed
  22. Quintela M, et al. 2023. Br J Cancer. . PubMed
  23. McKenzie C, et al. 2023. Mol Ther Nucleic Acids. 32:603. PubMed
  24. Lindner J, et al. 2013. Mol Cell Biol. 33:4628. PubMed
  25. Turner J, et al. 2013. J Exp Med. 210:2951. PubMed
  26. Itkin T, et al. 2016. Nature. 532: 323-328. PubMed
  27. Hergott C, et al. 2016. Blood. 127: 2460 - 2471. PubMed
  28. Wang J, et al. 2021. ACS Nano. 15:14360. PubMed
  29. Svoronos AA, et al. 2021. iScience. 24:103208. PubMed
  30. Wang Q, et al. 2022. Nat Commun. 13:3022. PubMed
  31. Olelewe C, et al. 2022. iScience. 25:104340. PubMed
  32. Lim JA, et al. 2018. BMC Cancer. 0.860416667. PubMed
  33. Venter C, et al. 2017. Cell Physiol Biochem. 43:1288. PubMed
  34. Terashima S, et al. 2017. J Radiat Res. 10.1093/jrr/rrx025. PubMed
  35. Kim Y, et al. 2020. J Neurochem. 155:177. PubMed
  36. Liu Y, et al. 2022. Cancer Discov. :. PubMed
  37. Wang M, et al. 2022. Immun Inflamm Dis. 10:e626. PubMed
  38. McNeal AS, et al. 2021. Elife. 10:. PubMed
  39. Soprano M, et al. 2016. PLoS One. 11: 0155970. PubMed
  40. Li L, et al. 2016. Cell Death Dis. 7:e2439. PubMed
  41. Moose DL, et al. 2020. Cell Rep. 30:3864. PubMed
  42. Wurzer H, et al. 2021. Front Immunol. 12:619069. PubMed
  43. Figueroa-Valdés AI, et al. 2021. Front Bioeng Biotechnol. 9:619930. PubMed
  44. Yoshioka H, et al. 2021. Front Cell Dev Biol. 9:621045. PubMed
  45. Dregalla RC, et al. 2021. Stem Cell Res Ther. 12:547. PubMed
  46. Sun V, et al. 2021. Front Immunol. 12:716661. PubMed
  47. Herman KD, et al. 2022. Front Immunol. 13:956991. PubMed
  48. Coray M, et al. 2022. Int J Mol Sci. 23:. PubMed
  49. Vries M, et al. 2014. Am J Physiol Lung Cell Mol Physiol. 307:240. PubMed
  50. Naaman H,et al. 2017. PLoS One. 10.1371/journal.pone.0187077. PubMed
  51. Elakad O, et al. 2022. NPJ Precis Oncol. 6:52. PubMed
  52. Deng J, et al. 2021. Cell Death Dis. 12:978. PubMed
  53. Tian AL, et al. 2021. Cell Death Dis. 12:6. PubMed
  54. Hickman O, et al. 2016. Br J Cancer. 10.1038/bjc.2016.91. PubMed
  55. Hagan AS, et al. 2020. Development. 147:00:00. PubMed
  56. Feng S, et al. 2017. Cancer Sci. 108:12:00. PubMed
  57. Zhou L, et al. 2017. Tumour Biol. 39:1010428317694313. PubMed
  58. Kinsella S, et al. 2021. Cell Rep. 37:109789. PubMed
  59. Wilson AS, et al. 2022. Nat Commun. 13:528. PubMed
  60. Wu J, et al. 2020. Immunity. 53:115. PubMed
  61. Zhang S, et al. 2020. Front Immunol. 1.757638889. PubMed
  62. Li P, et al. 2016. Mol Biol Cell. 27: 2542 - 2553. PubMed
  63. Luca E, et al. 2020. Mol Metab. 36:100967. PubMed
  64. Saint Fleur–Lominy S, et al. 2018. Cell Rep. 24:3045. PubMed
  65. Krishnasamy K,et al. 2017. Nat Commun. . 10.1038/s41467-017-00953-2. PubMed
  66. Yang L, et al. 2021. Cell Death Differ. 28:2616. PubMed
  67. Burbulla L, et al. 2014. Cell Death Dis. 5:1180. PubMed
  68. Peiris T, et al. 2016. Development. 143: 1697 - 1709.. PubMed
  69. Herold N, et al. 2017. Nat Med. 23:256-263. PubMed
  70. Domenis R, et al. 2017. PLoS One. 10.1371/journal.pone.0169932. PubMed
  71. Xie S, et al. 2021. Sci Rep. 0.660416667. PubMed
  72. Hönes J, et al. 2017. Sci Rep. . 10.1038/s41598-017-15866-9. PubMed
  73. Wang P, et al. 2019. Nat Commun. 0.940972222. PubMed
  74. Celus W, et al. 2021. Cancer Immunol Res. Online ahead of print. PubMed
  75. Cheng CC, et al. 2020. Elife. 9:00. PubMed
  76. Leiendecker L, et al. 2020. EMBO Mol Med. 12:e12525. PubMed
  77. Jung E, et al. 2017. J Neurosci. 37:6837. PubMed
  78. Matsuda K, et al. 2021. Cell Rep Methods. 1:100122. PubMed
  79. Carosi JM, et al. 2021. Autophagy. 17:2217. PubMed
  80. Li Y, et al. 2021. Sci Transl Med. 13:. PubMed
  81. Thakran S, et al. 2015. Invest Ophthalmol Vis Sci . 56:177. PubMed
  82. Suarez S, et al. 2015. J Biol Chem. 290: 28311 - 28320. PubMed
  83. Deventhiran J, et al. 2016. J Virol. 90: 222 - 231. PubMed
  84. Verma N, et al. 2020. Sci Adv. 6:00. PubMed
  85. Litzenburger UM, et al. 2017. Genome Biol. 18:15. PubMed
  86. Bartkowiak K, et al. 2021. Cancers (Basel). 13: . PubMed
  87. Pedicona F, et al. 2022. Sci Signal. 15:eabl7989. PubMed
  88. Li C, et al. 2021. Cell Metabolism. 33(8):1610-1623.e5. PubMed
  89. L Chang, Y Li, D Kaplan 2016. Sci Rep. 6:36862. PubMed
  90. Becker A, et al. 2016. Sci Rep. 6: 33847. PubMed
  91. Hopkins T, et al. 2015. Nucleic Acids Res. 10.1093/nar/gkv1515. PubMed
  92. Bersini S, et al. 2020. Life Sci Alliance. 3:e201900623. PubMed
  93. Lasrado N, et al. 2022. iScience. 25:103865. PubMed
  94. Yabas M, et al. 2016. PLoS One. 11: 0146774. PubMed
  95. Li B, et al. 2019. Oncol Rep. 41:608. PubMed
  96. Wang M, et al. 2018. Diabetologia. 62:335. PubMed
  97. Textor A, et al. 2021. Cancers (Basel). 13:. PubMed
  98. Molinar-Inglis O, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  99. Erdogan M, et al. 2022. Chem Biodivers. 19:e202200659. PubMed
  100. Harro CM, et al. 2020. J Clin Invest. . PubMed
  101. Cabet E, et al. 2015. PLoS One. 10: 0137009. PubMed
  102. Hasselluhn MC, et al. 2019. Cell Death Dis. 0.726388889. PubMed
  103. Zhang S, et al. 2018. Sci Rep. 8:17066. PubMed
  104. Lee HW, et al. 2018. Am J Physiol Renal Physiol. 314:F280. PubMed
  105. Pascual-Pasto G, et al. 2022. J Immunother Cancer. 10:. PubMed
  106. Soon P, et al. 2013. Endocr Relat Cancer. 20:1. PubMed
  107. Dong G, et al. 2020. Haematologica. . PubMed
  108. Abu El Maaty MA, et al. 2021. Sci Adv. 7: . PubMed
  109. Bahl S, et al. 2021. Cell Death Dis. 12:469. PubMed
  110. Labi V, et al. 2019. Genes Dev. 33:1673. PubMed
  111. Sun T, et al. 2019. Cancer Med. 2.457638889. PubMed
  112. Kiss M, et al. 2020. Cancer Immunol Res. 9:309. PubMed
  113. Su PL, et al. 2021. Lung Cancer. 158:137. PubMed
  114. Beggs K, et al. 2014. Toxicol Sci. 137:91. PubMed
  115. Guentsch A, et al. 2017. Mol Cell Biol. 37: e00236-16. PubMed
  116. Festag J, et al. 2020. Mol Ther Nucleic Acids. 1.330555556. PubMed
  117. Marton N, et al. 2017. Cellular and Molecular Life Sciences. 10.1007/s00018-017-2535-8. PubMed
  118. Kranz P, et al. 2017. Cell Death Dis. 8:e2986. PubMed
  119. Wu J, et al. 2021. Adv Sci (Weinh). 8:e2101029. PubMed
  120. Kojima N, et al. 2015. Anticancer Res. 35: 4493-4499. PubMed
  121. Xiong R, et al. 2014. Toxicol Appl Pharmacol. 280:285. PubMed
  122. Pedro L, et al. 2019. Small. 15:e1900224. PubMed
  123. Rosenberger L, et al. 2019. Sci Rep. 0.835416667. PubMed
  124. Bensberg M, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  125. Sahin E, et al. 2014. J Immunol. 193:1717. PubMed
  126. Kim J, et al. 2019. PLoS One. 14:e0210248. PubMed
  127. Quan M, et al. 2019. Oncol Rep. 41:3051. PubMed
  128. Di Franco S, et al. 2021. STAR Protoc. 2:100880. PubMed
  129. Jatho A, et al. 2022. Cells. 11:. PubMed
  130. Langsch S, et al. 2016. Cancer Res . 76: 4160 - 4169. PubMed
  131. Xiong R, et al. 2015. Mol Pharmacol. 88: 1045 - 1054. PubMed
  132. Vancsik T, et al. 2019. Cancer Med. 3.313888889. PubMed
  133. Vulin M, et al. 2022. Oncogene. :. PubMed
  134. Guo P, et al. 2022. Nat Cell Biol. 24:99. 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 or therapeutic use.

 

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