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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 CHF
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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 (pdf)

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. Shrestha C,et al. 2017. PLoS One. 10.1371/journal.pone.0186169. PubMed
  2. Djeungoue–Petga MA, et al. 2019. Cell Death Dis. 1.069444444. PubMed
  3. Wang YH, et al. 2022. Sci Adv. 8:eabn6552. PubMed
  4. Vajda F, et al. 2022. Cells. 11:. PubMed
  5. Chu CY, et al. 2023. Sci Rep. 13:3943. PubMed
  6. Riera-Domingo C, et al. 2023. Sci Adv. 9:eadd5028. PubMed
  7. Bujarrabal-Dueso A, et al. 2023. Nat Struct Mol Biol. 30:475. PubMed
  8. Quintela M, et al. 2023. Clin Epigenetics. 15:63. PubMed
  9. Quintela M, et al. 2023. Br J Cancer. . PubMed
  10. McKenzie C, et al. 2023. Mol Ther Nucleic Acids. 32:603. PubMed
  11. Forika G, et al. 2021. Pathol Oncol Res. 27:1610048. PubMed
  12. Chen Q, et al. 2022. Cell Rep. 39:110990. PubMed
  13. Clusmann J, et al. 2022. Cell Death Dis. 13:702. PubMed
  14. Hao J, et al. 2022. Cell Rep. 41:111804. PubMed
  15. Shahid AM, et al. 2022. PLoS One. 17:e0278209. PubMed
  16. Cappellesso F, et al. 2022. Nat Cancer. 3:1464. PubMed
  17. Muckenhuber M, et al. 2023. Am J Transplant. 23:84. PubMed
  18. Zhao Q, et al. 2023. Nat Commun. 14:735. PubMed
  19. Tsuji M 2014. Mol Biol Cell. 25:2116. PubMed
  20. Guo H, et al. 2016. Sci Rep. 6:37559. PubMed
  21. Kitazawa M, et al. 2017. PLoS One. 10.1371/journal.pone.0169340. PubMed
  22. Liu M, et al. 2021. Mol Med Rep. 23:00. PubMed
  23. Ocklenburg T, et al. 2021. Sci Rep. 7199:11. PubMed
  24. Wang J, et al. 2021. ACS Nano. 15:14360. PubMed
  25. Svoronos AA, et al. 2021. iScience. 24:103208. PubMed
  26. Wang Q, et al. 2022. Nat Commun. 13:3022. PubMed
  27. Olelewe C, et al. 2022. iScience. 25:104340. PubMed
  28. Lindner J, et al. 2013. Mol Cell Biol. 33:4628. PubMed
  29. Turner J, et al. 2013. J Exp Med. 210:2951. PubMed
  30. Itkin T, et al. 2016. Nature. 532: 323-328. PubMed
  31. Hergott C, et al. 2016. Blood. 127: 2460 - 2471. 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. Moose DL, et al. 2020. Cell Rep. 30:3864. PubMed
  40. Wurzer H, et al. 2021. Front Immunol. 12:619069. PubMed
  41. Figueroa-Valdés AI, et al. 2021. Front Bioeng Biotechnol. 9:619930. PubMed
  42. Yoshioka H, et al. 2021. Front Cell Dev Biol. 9:621045. PubMed
  43. Dregalla RC, et al. 2021. Stem Cell Res Ther. 12:547. PubMed
  44. Sun V, et al. 2021. Front Immunol. 12:716661. PubMed
  45. Herman KD, et al. 2022. Front Immunol. 13:956991. PubMed
  46. Coray M, et al. 2022. Int J Mol Sci. 23:. PubMed
  47. Soprano M, et al. 2016. PLoS One. 11: 0155970. PubMed
  48. Li L, et al. 2016. Cell Death Dis. 7:e2439. PubMed
  49. Naaman H,et al. 2017. PLoS One. 10.1371/journal.pone.0187077. PubMed
  50. Elakad O, et al. 2022. NPJ Precis Oncol. 6:52. PubMed
  51. Deng J, et al. 2021. Cell Death Dis. 12:978. PubMed
  52. Tian AL, et al. 2021. Cell Death Dis. 12:6. PubMed
  53. Vries M, et al. 2014. Am J Physiol Lung Cell Mol Physiol. 307:240. PubMed
  54. Feng S, et al. 2017. Cancer Sci. 108:12:00. PubMed
  55. Zhou L, et al. 2017. Tumour Biol. 39:1010428317694313. PubMed
  56. Kinsella S, et al. 2021. Cell Rep. 37:109789. PubMed
  57. Wilson AS, et al. 2022. Nat Commun. 13:528. PubMed
  58. Hickman O, et al. 2016. Br J Cancer. 10.1038/bjc.2016.91. PubMed
  59. Hagan AS, et al. 2020. Development. 147:00:00. PubMed
  60. Luca E, et al. 2020. Mol Metab. 36:100967. PubMed
  61. Saint Fleur–Lominy S, et al. 2018. Cell Rep. 24:3045. PubMed
  62. Krishnasamy K,et al. 2017. Nat Commun. . 10.1038/s41467-017-00953-2. PubMed
  63. Yang L, et al. 2021. Cell Death Differ. 28:2616. PubMed
  64. Wu J, et al. 2020. Immunity. 53:115. PubMed
  65. Zhang S, et al. 2020. Front Immunol. 1.757638889. PubMed
  66. Li P, et al. 2016. Mol Biol Cell. 27: 2542 - 2553. PubMed
  67. Hönes J, et al. 2017. Sci Rep. . 10.1038/s41598-017-15866-9. PubMed
  68. Wang P, et al. 2019. Nat Commun. 0.940972222. PubMed
  69. Celus W, et al. 2021. Cancer Immunol Res. Online ahead of print. PubMed
  70. Burbulla L, et al. 2014. Cell Death Dis. 5:1180. PubMed
  71. Peiris T, et al. 2016. Development. 143: 1697 - 1709.. PubMed
  72. Herold N, et al. 2017. Nat Med. 23:256-263. PubMed
  73. Domenis R, et al. 2017. PLoS One. 10.1371/journal.pone.0169932. PubMed
  74. Xie S, et al. 2021. Sci Rep. 0.660416667. PubMed
  75. Jung E, et al. 2017. J Neurosci. 37:6837. PubMed
  76. Matsuda K, et al. 2021. Cell Rep Methods. 1:100122. PubMed
  77. Carosi JM, et al. 2021. Autophagy. 17:2217. PubMed
  78. Li Y, et al. 2021. Sci Transl Med. 13:. PubMed
  79. Cheng CC, et al. 2020. Elife. 9:00. PubMed
  80. Leiendecker L, et al. 2020. EMBO Mol Med. 12:e12525. PubMed
  81. Litzenburger UM, et al. 2017. Genome Biol. 18:15. PubMed
  82. Bartkowiak K, et al. 2021. Cancers (Basel). 13: . PubMed
  83. Pedicona F, et al. 2022. Sci Signal. 15:eabl7989. PubMed
  84. Thakran S, et al. 2015. Invest Ophthalmol Vis Sci . 56:177. PubMed
  85. Suarez S, et al. 2015. J Biol Chem. 290: 28311 - 28320. PubMed
  86. Deventhiran J, et al. 2016. J Virol. 90: 222 - 231. PubMed
  87. Verma N, et al. 2020. Sci Adv. 6:00. PubMed
  88. Bersini S, et al. 2020. Life Sci Alliance. 3:e201900623. PubMed
  89. Lasrado N, et al. 2022. iScience. 25:103865. PubMed
  90. Li C, et al. 2021. Cell Metabolism. 33(8):1610-1623.e5. PubMed
  91. L Chang, Y Li, D Kaplan 2016. Sci Rep. 6:36862. PubMed
  92. Becker A, et al. 2016. Sci Rep. 6: 33847. PubMed
  93. Hopkins T, et al. 2015. Nucleic Acids Res. 10.1093/nar/gkv1515. PubMed
  94. Li B, et al. 2019. Oncol Rep. 41:608. PubMed
  95. Wang M, et al. 2018. Diabetologia. 62:335. PubMed
  96. Textor A, et al. 2021. Cancers (Basel). 13:. PubMed
  97. Molinar-Inglis O, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  98. Erdogan M, et al. 2022. Chem Biodivers. 19:e202200659. PubMed
  99. Yabas M, et al. 2016. PLoS One. 11: 0146774. PubMed
  100. Hasselluhn MC, et al. 2019. Cell Death Dis. 0.726388889. PubMed
  101. Zhang S, et al. 2018. Sci Rep. 8:17066. PubMed
  102. Lee HW, et al. 2018. Am J Physiol Renal Physiol. 314:F280. PubMed
  103. Pascual-Pasto G, et al. 2022. J Immunother Cancer. 10:. PubMed
  104. Harro CM, et al. 2020. J Clin Invest. . PubMed
  105. Cabet E, et al. 2015. PLoS One. 10: 0137009. PubMed
  106. Abu El Maaty MA, et al. 2021. Sci Adv. 7: . PubMed
  107. Bahl S, et al. 2021. Cell Death Dis. 12:469. PubMed
  108. Soon P, et al. 2013. Endocr Relat Cancer. 20:1. PubMed
  109. Dong G, et al. 2020. Haematologica. . 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. Marton N, et al. 2017. Cellular and Molecular Life Sciences. 10.1007/s00018-017-2535-8. PubMed
  115. Kranz P, et al. 2017. Cell Death Dis. 8:e2986. PubMed
  116. Wu J, et al. 2021. Adv Sci (Weinh). 8:e2101029. PubMed
  117. Beggs K, et al. 2014. Toxicol Sci. 137:91. PubMed
  118. Guentsch A, et al. 2017. Mol Cell Biol. 37: e00236-16. PubMed
  119. Festag J, et al. 2020. Mol Ther Nucleic Acids. 1.330555556. PubMed
  120. Pedro L, et al. 2019. Small. 15:e1900224. PubMed
  121. Rosenberger L, et al. 2019. Sci Rep. 0.835416667. PubMed
  122. Bensberg M, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  123. Kojima N, et al. 2015. Anticancer Res. 35: 4493-4499. PubMed
  124. Xiong R, et al. 2014. Toxicol Appl Pharmacol. 280:285. PubMed
  125. Kim J, et al. 2019. PLoS One. 14:e0210248. PubMed
  126. Quan M, et al. 2019. Oncol Rep. 41:3051. PubMed
  127. Di Franco S, et al. 2021. STAR Protoc. 2:100880. PubMed
  128. Jatho A, et al. 2022. Cells. 11:. PubMed
  129. Sahin E, et al. 2014. J Immunol. 193:1717. PubMed
  130. Vancsik T, et al. 2019. Cancer Med. 3.313888889. PubMed
  131. Vulin M, et al. 2022. Oncogene. :. PubMed
  132. Guo P, et al. 2022. Nat Cell Biol. 24:99. PubMed
  133. Langsch S, et al. 2016. Cancer Res . 76: 4160 - 4169. PubMed
  134. Xiong R, et al. 2015. Mol Pharmacol. 88: 1045 - 1054. PubMed

Antigen Details

Gene ID
NA
Specificity (DOES NOT SHOW ON TDS):
Annexin V
Specificity Alt (DOES NOT SHOW ON TDS):
Annexin V
App Abbreviation (DOES NOT SHOW ON TDS):
FC

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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