Loading...

Alexa Fluor® 647 Annexin V

Pricing & Availability
Regulatory Status
RUO
Other Names
Annexin A5
Ave. Rating
Submit a Review
Product Citations
publications
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
Cat # Size Price Quantity Check Availability Save
640911 25 tests 81€
Check Availability


Need larger quantities of this item?
Request Bulk Quote
640912 100 tests 188€
Check Availability


Need larger quantities of this item?
Request Bulk Quote
640943 300 tests 372€
Check Availability


Need larger quantities of this item?
Request Bulk Quote
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 (pdf)

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.

View full statement regarding label licenses
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
  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. Forika G, et al. 2021. Pathol Oncol Res. 27:1610048. PubMed
  2. Manzano-Muñoz A, et al. 2022. NPJ Precis Oncol. 6:90. PubMed
  3. Berastegui N, et al. 2022. Nat Commun. 13:7619. PubMed
  4. Pinci F, et al. 2022. Front Immunol. 13:1074440. PubMed
  5. Kitazawa M, et al. 2017. PLoS One. 10.1371/journal.pone.0169340. PubMed
  6. Berthenet K, et al. 2020. Cell Rep. 31:107731. PubMed
  7. Li W, et al. 2019. Aging (Albany NY). 0.99375. PubMed
  8. Rashid R, et al. 2019. Sci Data. 0.474305556. 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. Csomós I, et al. 2021. Int J Mol Sci. 22:. PubMed
  24. Leib P 2012. MBio. 16:267. PubMed
  25. Abaeva A, et al. 2013. J Biol Chem. 288:29621. PubMed
  26. Alcon C, et al. 2020. Cell Death Dis. 0.898611111. PubMed
  27. Hug N, et al. 2022. RNA. 28:1224. PubMed
  28. Hinterbrandner M, et al. 2021. JCI Insight. 6:e151797. PubMed
  29. Hirano KI, et al. 2021. eLife. 0.416666666666667. PubMed
  30. Schlicher L, et al. 2016. EMBO Rep. 17: 1485 - 1497. PubMed
  31. Xu L, et al. 2012. J Immunol. 189:3347. PubMed
  32. Wurzer H, et al. 2021. Front Immunol. 12:619069. PubMed
  33. Rashad S, et al. 2021. J Cell Physiol. 236:3710. PubMed
  34. Ohtsuka S, et al. 2016. Int Immunol. 28: 547 - 557. PubMed
  35. He Y, et al. 2020. Nat Commun. 11:1996. PubMed
  36. Tian AL, et al. 2021. Cell Death Dis. 12:6. PubMed
  37. Lopez J, et al. 2016. Nat Commun. 7:10538. PubMed
  38. Zakharova N, et al. 2015. PLoS One. 10:116665. PubMed
  39. Wortmann A, et al. 2011. J Biol Chem. 286:42303. PubMed
  40. Wu X, et al. 2020. Front Biosci (Landmark Ed). 25:513. PubMed
  41. Schögler A, et al. 2015. Eur Respir J. 45:428. PubMed
  42. Artemenko E, et al. 2016. Biochem J. 473: 435 - 448. PubMed
  43. Lin JR et al. 2018. eLife. 7 pii: e31657. 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. Carosi JM, et al. 2021. Autophagy. 17:2217. PubMed
  51. Smith CM, et al. 2021. Biochem J. 478:3331. PubMed
  52. Zhang X, et al. 2020. Eur J Med Chem. 112186:192. PubMed
  53. Yue D, et al. 2015. Exp Cell Res. 336: 141-149. PubMed
  54. Fan Z, et al. 2019. Aging (Albany NY). 0.85625. 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. Pal P, et al. 2021. J Med Chem. 64:14230. PubMed
  67. Kinoshita M, et al. 2021. Proc Natl Acad Sci U S A. 118:. PubMed
  68. Canedo EC, et al. 2021. JCI Insight. 6:. PubMed
  69. Basu T, et al. 2021. Hemasphere. 5:e565. PubMed
  70. Adams O, et al. 2018. PLoS One. 13:e0197610. PubMed
  71. Manzano-Muñoz A, et al. 2021. Front Cell Dev Biol. 9:695225. PubMed
  72. Bobylev I, et al. 2017. Neurotox Res.. 10.1007/s12640-017-9760-7. PubMed
  73. Mendelson A, et al. 2019. JCI Insight. 4:e126982. PubMed
  74. Zimmerman M, et al. 2010. PLoS One. 5:e14076. PubMed
  75. Alcon C, et al. 2021. Cells. 10:. PubMed
  76. Podoplelova N, et al. 2016. Blood. 128: 1745 - 1755. PubMed
  77. Bao W, et al. 2016. Sci Rep. 6:22579. PubMed
  78. Ghura H, et al. 2021. Neoplasia. 23:837. PubMed
  79. Liu F, et al. 2012. J Biol Chem. 287:25530. PubMed
  80. Beggs K, et al. 2014. Toxicol Sci. 137:91. PubMed
  81. Akhtar N, et al. 2016. Dev Cell. 38: 522-535. PubMed
  82. Yang D, et al. 2012. J Immunol. 188:4441. 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. Jatho A, et al. 2022. Cells. 11:. PubMed
  90. Mauro AN, et al. 2022. Sci Rep. 12:14537. PubMed
  91. Topalov N, et al. 2012. Arterioscler Thromb Vasc Biol. 32:2475. PubMed
  92. Ponzetta A, et al. 2013. J Immunol. 191:5684. PubMed
  93. K?fer R, et al. 2017. Molecular Immunology. 10.1016/j.molimm.2017.05.003. PubMed
  94. Lima PDA, et al. 2018. Biol Reprod. 99:838. PubMed
  95. Ho J, et al. 2011. PLoS One. 6:e16815. PubMed
  96. Vulin M, et al. 2022. Oncogene. :. PubMed
  97. Duell J, et al. 2017. Leukemia. 10.1038/leu.2017.41. PubMed
  98. Schmid M, et al. 2012. Immunobiology. 217:610. PubMed
  99. Zammarchi F, et al. 2011. Proc Natl Acad Sci U S A. 108:17779. PubMed
  100. Zeevi D, et al. 2010. J Cell Sci. 123:3112. PubMed
  101. Santidrián A, et al. 2010. Blood. 116:3023. PubMed
  102. Lin PB, et al. 2019. Oncol Rep. 41:3377. 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.

 

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