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 Quantity Check Availability Save
422201 100 mL 29€
Check Availability


Need larger quantities of this item?
Request Bulk Quote
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. 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. Lindner J, et al. 2013. Mol Cell Biol. 33:4628. PubMed
  10. Turner J, et al. 2013. J Exp Med. 210:2951. PubMed
  11. Itkin T, et al. 2016. Nature. 532: 323-328. PubMed
  12. Hergott C, et al. 2016. Blood. 127: 2460 - 2471. PubMed
  13. Wang J, et al. 2021. ACS Nano. 15:14360. PubMed
  14. Svoronos AA, et al. 2021. iScience. 24:103208. PubMed
  15. Wang Q, et al. 2022. Nat Commun. 13:3022. PubMed
  16. Olelewe C, et al. 2022. iScience. 25:104340. PubMed
  17. Lim JA, et al. 2018. BMC Cancer. 0.860416667. PubMed
  18. Venter C, et al. 2017. Cell Physiol Biochem. 43:1288. PubMed
  19. Terashima S, et al. 2017. J Radiat Res. 10.1093/jrr/rrx025. PubMed
  20. Kim Y, et al. 2020. J Neurochem. 155:177. PubMed
  21. Liu Y, et al. 2022. Cancer Discov. :. PubMed
  22. Wang M, et al. 2022. Immun Inflamm Dis. 10:e626. PubMed
  23. McNeal AS, et al. 2021. Elife. 10:. PubMed
  24. Soprano M, et al. 2016. PLoS One. 11: 0155970. PubMed
  25. Li L, et al. 2016. Cell Death Dis. 7:e2439. PubMed
  26. Moose DL, et al. 2020. Cell Rep. 30:3864. PubMed
  27. Wurzer H, et al. 2021. Front Immunol. 12:619069. PubMed
  28. Figueroa-Valdés AI, et al. 2021. Front Bioeng Biotechnol. 9:619930. PubMed
  29. Yoshioka H, et al. 2021. Front Cell Dev Biol. 9:621045. PubMed
  30. Dregalla RC, et al. 2021. Stem Cell Res Ther. 12:547. PubMed
  31. Sun V, et al. 2021. Front Immunol. 12:716661. PubMed
  32. Herman KD, et al. 2022. Front Immunol. 13:956991. PubMed
  33. Coray M, et al. 2022. Int J Mol Sci. 23:. PubMed
  34. Vries M, et al. 2014. Am J Physiol Lung Cell Mol Physiol. 307:240. PubMed
  35. Naaman H,et al. 2017. PLoS One. 10.1371/journal.pone.0187077. PubMed
  36. Elakad O, et al. 2022. NPJ Precis Oncol. 6:52. PubMed
  37. Deng J, et al. 2021. Cell Death Dis. 12:978. PubMed
  38. Tian AL, et al. 2021. Cell Death Dis. 12:6. PubMed
  39. Hickman O, et al. 2016. Br J Cancer. 10.1038/bjc.2016.91. PubMed
  40. Hagan AS, et al. 2020. Development. 147:00:00. PubMed
  41. Feng S, et al. 2017. Cancer Sci. 108:12:00. PubMed
  42. Zhou L, et al. 2017. Tumour Biol. 39:1010428317694313. PubMed
  43. Kinsella S, et al. 2021. Cell Rep. 37:109789. PubMed
  44. Wilson AS, et al. 2022. Nat Commun. 13:528. PubMed
  45. Wu J, et al. 2020. Immunity. 53:115. PubMed
  46. Zhang S, et al. 2020. Front Immunol. 1.757638889. PubMed
  47. Li P, et al. 2016. Mol Biol Cell. 27: 2542 - 2553. PubMed
  48. Luca E, et al. 2020. Mol Metab. 36:100967. PubMed
  49. Saint Fleur–Lominy S, et al. 2018. Cell Rep. 24:3045. PubMed
  50. Krishnasamy K,et al. 2017. Nat Commun. . 10.1038/s41467-017-00953-2. PubMed
  51. Yang L, et al. 2021. Cell Death Differ. 28:2616. PubMed
  52. Burbulla L, et al. 2014. Cell Death Dis. 5:1180. PubMed
  53. Peiris T, et al. 2016. Development. 143: 1697 - 1709.. PubMed
  54. Herold N, et al. 2017. Nat Med. 23:256-263. PubMed
  55. Domenis R, et al. 2017. PLoS One. 10.1371/journal.pone.0169932. PubMed
  56. Xie S, et al. 2021. Sci Rep. 0.660416667. PubMed
  57. Hönes J, et al. 2017. Sci Rep. . 10.1038/s41598-017-15866-9. PubMed
  58. Wang P, et al. 2019. Nat Commun. 0.940972222. PubMed
  59. Celus W, et al. 2021. Cancer Immunol Res. Online ahead of print. PubMed
  60. Cheng CC, et al. 2020. Elife. 9:00. PubMed
  61. Leiendecker L, et al. 2020. EMBO Mol Med. 12:e12525. PubMed
  62. Jung E, et al. 2017. J Neurosci. 37:6837. PubMed
  63. Matsuda K, et al. 2021. Cell Rep Methods. 1:100122. PubMed
  64. Carosi JM, et al. 2021. Autophagy. 17:2217. PubMed
  65. Li Y, et al. 2021. Sci Transl Med. 13:. PubMed
  66. Thakran S, et al. 2015. Invest Ophthalmol Vis Sci . 56:177. PubMed
  67. Suarez S, et al. 2015. J Biol Chem. 290: 28311 - 28320. PubMed
  68. Deventhiran J, et al. 2016. J Virol. 90: 222 - 231. PubMed
  69. Verma N, et al. 2020. Sci Adv. 6:00. PubMed
  70. Litzenburger UM, et al. 2017. Genome Biol. 18:15. PubMed
  71. Bartkowiak K, et al. 2021. Cancers (Basel). 13: . PubMed
  72. Pedicona F, et al. 2022. Sci Signal. 15:eabl7989. PubMed
  73. Li C, et al. 2021. Cell Metabolism. 33(8):1610-1623.e5. PubMed
  74. L Chang, Y Li, D Kaplan 2016. Sci Rep. 6:36862. PubMed
  75. Becker A, et al. 2016. Sci Rep. 6: 33847. PubMed
  76. Hopkins T, et al. 2015. Nucleic Acids Res. 10.1093/nar/gkv1515. PubMed
  77. Bersini S, et al. 2020. Life Sci Alliance. 3:e201900623. PubMed
  78. Lasrado N, et al. 2022. iScience. 25:103865. PubMed
  79. Yabas M, et al. 2016. PLoS One. 11: 0146774. PubMed
  80. Li B, et al. 2019. Oncol Rep. 41:608. PubMed
  81. Wang M, et al. 2018. Diabetologia. 62:335. PubMed
  82. Textor A, et al. 2021. Cancers (Basel). 13:. PubMed
  83. Molinar-Inglis O, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  84. Erdogan M, et al. 2022. Chem Biodivers. 19:e202200659. PubMed
  85. Harro CM, et al. 2020. J Clin Invest. . PubMed
  86. Cabet E, et al. 2015. PLoS One. 10: 0137009. PubMed
  87. Hasselluhn MC, et al. 2019. Cell Death Dis. 0.726388889. PubMed
  88. Zhang S, et al. 2018. Sci Rep. 8:17066. PubMed
  89. Lee HW, et al. 2018. Am J Physiol Renal Physiol. 314:F280. PubMed
  90. Pascual-Pasto G, et al. 2022. J Immunother Cancer. 10:. PubMed
  91. Soon P, et al. 2013. Endocr Relat Cancer. 20:1. PubMed
  92. Dong G, et al. 2020. Haematologica. . PubMed
  93. Abu El Maaty MA, et al. 2021. Sci Adv. 7: . PubMed
  94. Bahl S, et al. 2021. Cell Death Dis. 12:469. PubMed
  95. Labi V, et al. 2019. Genes Dev. 33:1673. PubMed
  96. Sun T, et al. 2019. Cancer Med. 2.457638889. PubMed
  97. Kiss M, et al. 2020. Cancer Immunol Res. 9:309. PubMed
  98. Su PL, et al. 2021. Lung Cancer. 158:137. PubMed
  99. Beggs K, et al. 2014. Toxicol Sci. 137:91. PubMed
  100. Guentsch A, et al. 2017. Mol Cell Biol. 37: e00236-16. PubMed
  101. Festag J, et al. 2020. Mol Ther Nucleic Acids. 1.330555556. PubMed
  102. Marton N, et al. 2017. Cellular and Molecular Life Sciences. 10.1007/s00018-017-2535-8. PubMed
  103. Kranz P, et al. 2017. Cell Death Dis. 8:e2986. PubMed
  104. Wu J, et al. 2021. Adv Sci (Weinh). 8:e2101029. PubMed
  105. Kojima N, et al. 2015. Anticancer Res. 35: 4493-4499. PubMed
  106. Xiong R, et al. 2014. Toxicol Appl Pharmacol. 280:285. PubMed
  107. Pedro L, et al. 2019. Small. 15:e1900224. PubMed
  108. Rosenberger L, et al. 2019. Sci Rep. 0.835416667. PubMed
  109. Bensberg M, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  110. Sahin E, et al. 2014. J Immunol. 193:1717. PubMed
  111. Kim J, et al. 2019. PLoS One. 14:e0210248. PubMed
  112. Quan M, et al. 2019. Oncol Rep. 41:3051. PubMed
  113. Di Franco S, et al. 2021. STAR Protoc. 2:100880. PubMed
  114. Jatho A, et al. 2022. Cells. 11:. PubMed
  115. Langsch S, et al. 2016. Cancer Res . 76: 4160 - 4169. PubMed
  116. Xiong R, et al. 2015. Mol Pharmacol. 88: 1045 - 1054. PubMed
  117. Vancsik T, et al. 2019. Cancer Med. 3.313888889. PubMed
  118. Vulin M, et al. 2022. Oncogene. :. PubMed
  119. 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.

 

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