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Alexa Fluor® 647 Annexin V

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Regulatory Status
RUO
Other Names
Annexin A5
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A647_Annexin-V_kit_052318
Human T leukemia cell line Jurkat, treated (left) or non-treated (right) with BioLegend’s anti-human CD95 (EOS9.1) mAb (Cat. No. 305704) for 4 hours, then stained with Annexin V- Alexa Fluor® 647 and Helix NP Green (Cat. No. 425303 at 1.25 nM) in Annexin V Binding buffer for 15 minutes at at 37°C.
  • A647_Annexin-V_kit_052318
    Human T leukemia cell line Jurkat, treated (left) or non-treated (right) with BioLegend’s anti-human CD95 (EOS9.1) mAb (Cat. No. 305704) for 4 hours, then stained with Annexin V- Alexa Fluor® 647 and Helix NP Green (Cat. No. 425303 at 1.25 nM) in Annexin V Binding buffer for 15 minutes at at 37°C.
See Alexa Fluor® 647 spectral data
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640911 25 tests 81€
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640912 100 tests 188€
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640943 300 tests 372€
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Description

Annexin V (or Annexin A5) is a member of the annexin family of intracellular proteins that binds to phosphatidylserine (PS) in a calcium-dependent manner. PS is normally only found on the intracellular leaflet of the plasma membrane in healthy cells, but during early apoptosis, membrane asymmetry is lost and PS translocates to the external leaflet. Fluorochrome-labeled Annexin V can then be used to specifically target and identify apoptotic cells.Annexin V Binding Buffer (cat. no. 422201) is recommended for use with Annexin V staining.Annexin V binding alone cannot differentiate between apoptotic cells and necrotic. Therefore, we recommend using our Helix NP™ Blue (Cat. No. 425305), Helix NP™ Green (Cat. No. 425303) or Helix NP™ NIR (Cat. No. 425301). Early apoptotic cells will exclude 7-AAD and PI, while late stage apoptotic cells and necrotic cells will stain positively, due to the passage of these dyes into the nucleus where they bind to DNA.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse, Rat
Reported Reactivity
Other Species
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation
The purified protein was conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The Annexin V solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC - Quality tested
Recommended Usage

Each lot of this product is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µL per 100,000 - million cells in a 100 µL volume of Annexin V Binding Buffer (Cat No. 422201). It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
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. Koopman G, et al. 1994. Blood 84:1415.
  2. Vermes I, et al. 1995. J. Immunol. Methods 184:39.
  3. Dachary-Prigent J et al. 1993. Blood 81:2554.
  4. Lev S, et al. 2010. J. Biol Chem. 287:2771. PubMed
  5. Jin D, et al. 2010. Cancer Res. 70:2245. PubMed
  6. Santidrian AF, et al. 2010. Blood 116:3023. PubMed
  7. Zammarchi, F., et al. 2011. PNAS. 108:17779. PubMed
  8. Liu F, et al. 2011. Cancer Res. 71:6807. PubMed
  9. Wortmann A, et al. 2011. J Biol Chem. 286:42303. PubMed
  10. Yang D, et al. 2012. J Immunol. 188:4441. PubMed
  11. Notake T, et al. 2012. J Immunol. 188:4838. PubMed
  12. Liu F, et al. 2012. J. Biol Chem. 287:25530. PubMed
  13. Schmid M, et al. 2012. Immunobiology. 217:610. PubMed
  14. Topalov NN, et al. 2012. Arterioscler Thromb Vasc Biol. 32:2475. PubMed
  15. Xu LS, et al. 2012. J. Immunol. 189:3347. PubMed
  16. Gobeil PA, et al. 2012. MBio. 16:267. PubMed
  17. Abaeva AA, et al. 2013. J Biol Chem. 288:29621. PubMed
  18. Ponzetta A, et al. 2013. J. Immunol. 191:5684. PubMed
  19. Beggs KM, et al. 2014. Toxicol Sci. 137:91. PubMed
  20. Yue D, et al. 2014. Exp Cell Res. 322:149. PubMed
  21. Schott J, et al. 2014. PLoS Genet. 10:1004368. PubMed
  22. Yasunaga M, et al. 2014. Sci Rep. 4:4852. PubMed
  23. Schogler A, et al. 2015. Eur Respir J. 45:428. PubMed
  24. Zakharova NV, et al. 2015. PLoS One. 10:116665. PubMed
  25. Ghalei H, et al. 2015. J Cell Biol. 208:745. PubMed
Product Citations
  1. Kitazawa M, et al. 2017. PLoS One. 10.1371/journal.pone.0169340. PubMed
  2. Berthenet K, et al. 2020. Cell Rep. 31:107731. PubMed
  3. Li W, et al. 2019. Aging (Albany NY). 0.99375. PubMed
  4. Rashid R, et al. 2019. Sci Data. 0.474305556. PubMed
  5. Forika G, et al. 2021. Pathol Oncol Res. 27:1610048. PubMed
  6. Manzano-Muñoz A, et al. 2022. NPJ Precis Oncol. 6:90. PubMed
  7. Berastegui N, et al. 2022. Nat Commun. 13:7619. PubMed
  8. Pinci F, et al. 2022. Front Immunol. 13:1074440. PubMed
  9. Wang YH, et al. 2022. Sci Adv. 8:eabn6552. PubMed
  10. Wagner AR, et al. 2022. Elife. 11:. PubMed
  11. Reynolds MB, et al. 2023. Sci Adv. 9:eade8701. PubMed
  12. Dufour S, et al. 2023. Commun Biol. 6:207. PubMed
  13. Cannavicci A, et al. 2023. Int J Mol Sci. 24:. PubMed
  14. Harpur CM, et al. 2023. Clin Transl Immunology. 12:e1443. PubMed
  15. Whyte D, et al. 2023. Mol Oncol. . PubMed
  16. Capellmann S, et al. 2023. Front Immunol. 14:1154416. PubMed
  17. Liu JC, et al. 2023. Int J Mol Sci. 24:. PubMed
  18. Sarno J, et al. 2023. Nat Commun. 14:2935. PubMed
  19. Ma X, et al. 2023. Research (Wash D C). 6:0124. PubMed
  20. Herz J, et al. 2015. J Exp Med. 212: 1153 - 1169. PubMed
  21. Liu X, et al. 2018. Bioorg Med Chem. 26:3925. PubMed
  22. Kanagalingam T, et al. 2019. Immun Inflamm Dis. 0.369444444. PubMed
  23. Leib P 2012. MBio. 16:267. PubMed
  24. Abaeva A, et al. 2013. J Biol Chem. 288:29621. PubMed
  25. Alcon C, et al. 2020. Cell Death Dis. 0.898611111. PubMed
  26. Csomós I, et al. 2021. Int J Mol Sci. 22:. PubMed
  27. Hirano KI, et al. 2021. eLife. 0.416666666666667. PubMed
  28. Schlicher L, et al. 2016. EMBO Rep. 17: 1485 - 1497. PubMed
  29. Xu L, et al. 2012. J Immunol. 189:3347. PubMed
  30. Hug N, et al. 2022. RNA. 28:1224. PubMed
  31. Hinterbrandner M, et al. 2021. JCI Insight. 6:e151797. PubMed
  32. Ohtsuka S, et al. 2016. Int Immunol. 28: 547 - 557. PubMed
  33. He Y, et al. 2020. Nat Commun. 11:1996. PubMed
  34. Wurzer H, et al. 2021. Front Immunol. 12:619069. PubMed
  35. Rashad S, et al. 2021. J Cell Physiol. 236:3710. PubMed
  36. Lopez J, et al. 2016. Nat Commun. 7:10538. PubMed
  37. Zakharova N, et al. 2015. PLoS One. 10:116665. PubMed
  38. Wortmann A, et al. 2011. J Biol Chem. 286:42303. PubMed
  39. Tian AL, et al. 2021. Cell Death Dis. 12:6. PubMed
  40. Schögler A, et al. 2015. Eur Respir J. 45:428. PubMed
  41. Artemenko E, et al. 2016. Biochem J. 473: 435 - 448. PubMed
  42. Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed
  43. Wu X, et al. 2020. Front Biosci (Landmark Ed). 25:513. PubMed
  44. Yue D, et al. 2014. Exp Cell Res. 322:149. PubMed
  45. Park A, et al. 2016. PLoS One. 11: 0148998. PubMed
  46. Coe G, et al. 2016. Sci Rep. 6:30816. PubMed
  47. Wiedemann GM, et al. 2020. Cell Rep. 33:108498. PubMed
  48. Lev S, et al. 2010. J Biol Chem. 285:2771. PubMed
  49. Notake T, et al. 2012. J Immunol. 188:4838. PubMed
  50. Zhang X, et al. 2020. Eur J Med Chem. 112186:192. PubMed
  51. Yue D, et al. 2015. Exp Cell Res. 336: 141-149. PubMed
  52. Fan Z, et al. 2019. Aging (Albany NY). 0.85625. PubMed
  53. Carosi JM, et al. 2021. Autophagy. 17:2217. PubMed
  54. Smith CM, et al. 2021. Biochem J. 478:3331. PubMed
  55. Schott J, et al. 2014. PLoS Genet. 10:1004368. PubMed
  56. Matsumura M 2014. Sci Rep. 4:4852. PubMed
  57. Ghalei H, et al. 2015. J Cell Biol. 208:745. PubMed
  58. Mair B, et al. 2016. PLoS Genet. 12: 1006279. PubMed
  59. Zhang Y, et al. 2020. Aging (Albany NY). 12:8067. PubMed
  60. Liu F, et al. 2011. Cancer Res. 71:6807. PubMed
  61. Jin D, et al. 2010. Cancer Res. 70:2245. PubMed
  62. Bersini S, et al. 2020. Life Sci Alliance. 3:e201900623. PubMed
  63. Hosokawa T, et al. 2017. J Immunol. 10.4049/jimmunol.1700157. PubMed
  64. Ma W, et al. 2017. Cell Death & Disease. 10.1038/cddis.2017.47. PubMed
  65. Rashad S, et al. 2020. RNA Biol. 1092:17. PubMed
  66. Adams O, et al. 2018. PLoS One. 13:e0197610. PubMed
  67. Pal P, et al. 2021. J Med Chem. 64:14230. PubMed
  68. Kinoshita M, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  69. Canedo EC, et al. 2021. JCI Insight. 6:. PubMed
  70. Basu T, et al. 2021. Hemasphere. 5:e565. PubMed
  71. Bobylev I, et al. 2017. Neurotox Res.. 10.1007/s12640-017-9760-7. PubMed
  72. Mendelson A, et al. 2019. JCI Insight. 4:e126982. PubMed
  73. Zimmerman M, et al. 2010. PLoS One. 5:e14076. PubMed
  74. Manzano-Muñoz A, et al. 2021. Front Cell Dev Biol. 9:695225. PubMed
  75. Podoplelova N, et al. 2016. Blood. 128: 1745 - 1755. PubMed
  76. Bao W, et al. 2016. Sci Rep. 6:22579. PubMed
  77. Alcon C, et al. 2021. Cells. 10:. PubMed
  78. Liu F, et al. 2012. J Biol Chem. 287:25530. PubMed
  79. Beggs K, et al. 2014. Toxicol Sci. 137:91. PubMed
  80. Akhtar N, et al. 2016. Dev Cell. 38: 522-535. PubMed
  81. Yang D, et al. 2012. J Immunol. 188:4441. PubMed
  82. Ghura H, et al. 2021. Neoplasia. 23:837. PubMed
  83. Gomez J, et al. 2016. J Immunol. 197: 2864 - 2879. PubMed
  84. Rossnagl S, et al. 2016. PLoS Biol. 14: 1002562. PubMed
  85. Camacho K, et al. 2015. J Control Release. 210: 198-207. PubMed
  86. He Y, et al. 2020. J Hematol Oncol. 0.607638889. PubMed
  87. Geary CD et al. 2018. Cell reports. 24(8):1949-1957 . PubMed
  88. Qiao Y,et al. 2017. Oncogene. . 10.1038/onc.2017.387. PubMed
  89. Topalov N, et al. 2012. Arterioscler Thromb Vasc Biol. 32:2475. PubMed
  90. Ponzetta A, et al. 2013. J Immunol. 191:5684. PubMed
  91. K?fer R, et al. 2017. Molecular Immunology. 10.1016/j.molimm.2017.05.003. PubMed
  92. Lima PDA, et al. 2018. Biol Reprod. 99:838. PubMed
  93. Ho J, et al. 2011. PLoS One. 6:e16815. PubMed
  94. Jatho A, et al. 2022. Cells. 11:. PubMed
  95. Mauro AN, et al. 2022. Sci Rep. 12:14537. PubMed
  96. Duell J, et al. 2017. Leukemia. 10.1038/leu.2017.41. PubMed
  97. Schmid M, et al. 2012. Immunobiology. 217:610. PubMed
  98. Zammarchi F, et al. 2011. Proc Natl Acad Sci U S A. 108:17779. PubMed
  99. Zeevi D, et al. 2010. J Cell Sci. 123:3112. PubMed
  100. Santidrián A, et al. 2010. Blood. 116:3023. PubMed
  101. Lin PB, et al. 2019. Oncol Rep. 41:3377. PubMed
  102. Vulin M, et al. 2022. Oncogene. :. PubMed
RRID
AB_2561293 (BioLegend Cat. No. 640911)
not an antibody (BioLegend Cat. No. 640912)
AB_2616658 (BioLegend Cat. No. 640943)

Antigen Details

Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Neuroscience
Gene ID
308 View all products for this Gene ID
UniProt
View information about Annexin V on UniProt.org

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: 7    Revision Date: 10.22.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

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