Loading...

Annexin V Binding Buffer

Pricing & Availability
Regulatory Status
RUO
Other Names
Annexin-V-Binding Buffer
Ave. Rating
Submit a Review
Product Citations
publications
Cat # Size Price Save
422201 100 mL ¥7,920
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. Forika G, et al. 2021. Pathol Oncol Res. 27:1610048. PubMed
  2. Chen Q, et al. 2022. Cell Rep. 39:110990. PubMed
  3. Clusmann J, et al. 2022. Cell Death Dis. 13:702. PubMed
  4. Hao J, et al. 2022. Cell Rep. 41:111804. PubMed
  5. Shahid AM, et al. 2022. PLoS One. 17:e0278209. PubMed
  6. Cappellesso F, et al. 2022. Nat Cancer. 3:1464. PubMed
  7. Muckenhuber M, et al. 2023. Am J Transplant. 23:84. PubMed
  8. Zhao Q, et al. 2023. Nat Commun. 14:735. PubMed
  9. Tsuji M 2014. Mol Biol Cell. 25:2116. PubMed
  10. Guo H, et al. 2016. Sci Rep. 6:37559. PubMed
  11. Kitazawa M, et al. 2017. PLoS One. 10.1371/journal.pone.0169340. PubMed
  12. Liu M, et al. 2021. Mol Med Rep. 23:00. PubMed
  13. Ocklenburg T, et al. 2021. Sci Rep. 7199:11. PubMed
  14. Wang YH, et al. 2022. Sci Adv. 8:eabn6552. PubMed
  15. Vajda F, et al. 2022. Cells. 11:. PubMed
  16. Chu CY, et al. 2023. Sci Rep. 13:3943. PubMed
  17. Riera-Domingo C, et al. 2023. Sci Adv. 9:eadd5028. PubMed
  18. Bujarrabal-Dueso A, et al. 2023. Nat Struct Mol Biol. 30:475. PubMed
  19. Quintela M, et al. 2023. Clin Epigenetics. 15:63. PubMed
  20. Quintela M, et al. 2023. Br J Cancer. . PubMed
  21. McKenzie C, et al. 2023. Mol Ther Nucleic Acids. 32:603. PubMed
  22. Shrestha C,et al. 2017. PLoS One. 10.1371/journal.pone.0186169. PubMed
  23. Djeungoue–Petga MA, et al. 2019. Cell Death Dis. 1.069444444. 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. Kim Y, et al. 2020. J Neurochem. 155:177. PubMed
  33. Liu Y, et al. 2022. Cancer Discov. :. PubMed
  34. Wang M, et al. 2022. Immun Inflamm Dis. 10:e626. PubMed
  35. McNeal AS, et al. 2021. Elife. 10:. PubMed
  36. Lim JA, et al. 2018. BMC Cancer. 0.860416667. PubMed
  37. Venter C, et al. 2017. Cell Physiol Biochem. 43:1288. PubMed
  38. Terashima S, et al. 2017. J Radiat Res. 10.1093/jrr/rrx025. PubMed
  39. Wurzer H, et al. 2021. Front Immunol. 12:619069. PubMed
  40. Figueroa-Valdés AI, et al. 2021. Front Bioeng Biotechnol. 9:619930. PubMed
  41. Yoshioka H, et al. 2021. Front Cell Dev Biol. 9:621045. PubMed
  42. Dregalla RC, et al. 2021. Stem Cell Res Ther. 12:547. PubMed
  43. Sun V, et al. 2021. Front Immunol. 12:716661. PubMed
  44. Herman KD, et al. 2022. Front Immunol. 13:956991. PubMed
  45. Coray M, et al. 2022. Int J Mol Sci. 23:. PubMed
  46. Soprano M, et al. 2016. PLoS One. 11: 0155970. PubMed
  47. Li L, et al. 2016. Cell Death Dis. 7:e2439. PubMed
  48. Moose DL, et al. 2020. Cell Rep. 30:3864. PubMed
  49. Elakad O, et al. 2022. NPJ Precis Oncol. 6:52. PubMed
  50. Deng J, et al. 2021. Cell Death Dis. 12:978. PubMed
  51. Tian AL, et al. 2021. Cell Death Dis. 12:6. PubMed
  52. Vries M, et al. 2014. Am J Physiol Lung Cell Mol Physiol. 307:240. PubMed
  53. Naaman H,et al. 2017. PLoS One. 10.1371/journal.pone.0187077. PubMed
  54. Kinsella S, et al. 2021. Cell Rep. 37:109789. PubMed
  55. Wilson AS, et al. 2022. Nat Commun. 13:528. PubMed
  56. Hickman O, et al. 2016. Br J Cancer. 10.1038/bjc.2016.91. PubMed
  57. Hagan AS, et al. 2020. Development. 147:00:00. PubMed
  58. Feng S, et al. 2017. Cancer Sci. 108:12:00. PubMed
  59. Zhou L, et al. 2017. Tumour Biol. 39:1010428317694313. PubMed
  60. Yang L, et al. 2021. Cell Death Differ. 28:2616. PubMed
  61. Wu J, et al. 2020. Immunity. 53:115. PubMed
  62. Zhang S, et al. 2020. Front Immunol. 1.757638889. PubMed
  63. Li P, et al. 2016. Mol Biol Cell. 27: 2542 - 2553. PubMed
  64. Luca E, et al. 2020. Mol Metab. 36:100967. PubMed
  65. Saint Fleur–Lominy S, et al. 2018. Cell Rep. 24:3045. PubMed
  66. Krishnasamy K,et al. 2017. Nat Commun. . 10.1038/s41467-017-00953-2. PubMed
  67. Celus W, et al. 2021. Cancer Immunol Res. Online ahead of print. PubMed
  68. Burbulla L, et al. 2014. Cell Death Dis. 5:1180. PubMed
  69. Peiris T, et al. 2016. Development. 143: 1697 - 1709.. PubMed
  70. Herold N, et al. 2017. Nat Med. 23:256-263. PubMed
  71. Domenis R, et al. 2017. PLoS One. 10.1371/journal.pone.0169932. PubMed
  72. Xie S, et al. 2021. Sci Rep. 0.660416667. PubMed
  73. Hönes J, et al. 2017. Sci Rep. . 10.1038/s41598-017-15866-9. PubMed
  74. Wang P, et al. 2019. Nat Commun. 0.940972222. PubMed
  75. Matsuda K, et al. 2021. Cell Rep Methods. 1:100122. PubMed
  76. Carosi JM, et al. 2021. Autophagy. 17:2217. PubMed
  77. Li Y, et al. 2021. Sci Transl Med. 13:. PubMed
  78. Cheng CC, et al. 2020. Elife. 9:00. PubMed
  79. Leiendecker L, et al. 2020. EMBO Mol Med. 12:e12525. PubMed
  80. Jung E, et al. 2017. J Neurosci. 37:6837. PubMed
  81. Bartkowiak K, et al. 2021. Cancers (Basel). 13: . PubMed
  82. Pedicona F, et al. 2022. Sci Signal. 15:eabl7989. PubMed
  83. Thakran S, et al. 2015. Invest Ophthalmol Vis Sci . 56:177. PubMed
  84. Suarez S, et al. 2015. J Biol Chem. 290: 28311 - 28320. PubMed
  85. Deventhiran J, et al. 2016. J Virol. 90: 222 - 231. PubMed
  86. Verma N, et al. 2020. Sci Adv. 6:00. PubMed
  87. Litzenburger UM, et al. 2017. Genome Biol. 18:15. PubMed
  88. Lasrado N, et al. 2022. iScience. 25:103865. PubMed
  89. Li C, et al. 2021. Cell Metabolism. 33(8):1610-1623.e5. PubMed
  90. L Chang, Y Li, D Kaplan 2016. Sci Rep. 6:36862. PubMed
  91. Becker A, et al. 2016. Sci Rep. 6: 33847. PubMed
  92. Hopkins T, et al. 2015. Nucleic Acids Res. 10.1093/nar/gkv1515. PubMed
  93. Bersini S, et al. 2020. Life Sci Alliance. 3:e201900623. PubMed
  94. Textor A, et al. 2021. Cancers (Basel). 13:. PubMed
  95. Molinar-Inglis O, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  96. Erdogan M, et al. 2022. Chem Biodivers. 19:e202200659. PubMed
  97. Yabas M, et al. 2016. PLoS One. 11: 0146774. PubMed
  98. Li B, et al. 2019. Oncol Rep. 41:608. PubMed
  99. Wang M, et al. 2018. Diabetologia. 62:335. PubMed
  100. Pascual-Pasto G, et al. 2022. J Immunother Cancer. 10:. PubMed
  101. Harro CM, et al. 2020. J Clin Invest. . PubMed
  102. Cabet E, et al. 2015. PLoS One. 10: 0137009. PubMed
  103. Hasselluhn MC, et al. 2019. Cell Death Dis. 0.726388889. PubMed
  104. Zhang S, et al. 2018. Sci Rep. 8:17066. PubMed
  105. Lee HW, et al. 2018. Am J Physiol Renal Physiol. 314:F280. 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. Kiss M, et al. 2020. Cancer Immunol Res. 9:309. PubMed
  111. Su PL, et al. 2021. Lung Cancer. 158:137. PubMed
  112. Labi V, et al. 2019. Genes Dev. 33:1673. PubMed
  113. Sun T, et al. 2019. Cancer Med. 2.457638889. PubMed
  114. Wu J, et al. 2021. Adv Sci (Weinh). 8:e2101029. PubMed
  115. Beggs K, et al. 2014. Toxicol Sci. 137:91. PubMed
  116. Guentsch A, et al. 2017. Mol Cell Biol. 37: e00236-16. PubMed
  117. Festag J, et al. 2020. Mol Ther Nucleic Acids. 1.330555556. PubMed
  118. Marton N, et al. 2017. Cellular and Molecular Life Sciences. 10.1007/s00018-017-2535-8. PubMed
  119. Kranz P, et al. 2017. Cell Death Dis. 8:e2986. PubMed
  120. Bensberg M, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  121. Kojima N, et al. 2015. Anticancer Res. 35: 4493-4499. PubMed
  122. Xiong R, et al. 2014. Toxicol Appl Pharmacol. 280:285. PubMed
  123. Pedro L, et al. 2019. Small. 15:e1900224. PubMed
  124. Rosenberger L, et al. 2019. Sci Rep. 0.835416667. PubMed
  125. Di Franco S, et al. 2021. STAR Protoc. 2:100880. PubMed
  126. Jatho A, et al. 2022. Cells. 11:. PubMed
  127. Sahin E, et al. 2014. J Immunol. 193:1717. PubMed
  128. Kim J, et al. 2019. PLoS One. 14:e0210248. PubMed
  129. Quan M, et al. 2019. Oncol Rep. 41:3051. PubMed
  130. Vulin M, et al. 2022. Oncogene. :. PubMed
  131. Guo P, et al. 2022. Nat Cell Biol. 24:99. PubMed
  132. Langsch S, et al. 2016. Cancer Res . 76: 4160 - 4169. PubMed
  133. Xiong R, et al. 2015. Mol Pharmacol. 88: 1045 - 1054. PubMed
  134. Vancsik T, et al. 2019. Cancer Med. 3.313888889. 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.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
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.

ProductsHere
Login / Register
Remember me
Forgot your password? Reset password?
Create an Account