Loading...

APC Annexin V

Pricing & Availability
Regulatory Status
RUO
Other Names
Annexin A5, Dead, Apoptosis
Ave. Rating
Submit a Review
Product Citations
publications
APC_Annexin-V_kit_052318_non-apop
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- APC and Helix NP Green (Cat. No. 425303 at 5 nM) in Annexin V Binding buffer for 15 minutes at 25°C.
  • APC_Annexin-V_kit_052318_non-apop
    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- APC and Helix NP Green (Cat. No. 425303 at 5 nM) in Annexin V Binding buffer for 15 minutes at 25°C.
See APC spectral data
Cat # Size Price Save
640919 25 tests ¥22,440
640920 100 tests ¥48,400
640941 300 tests ¥97,680
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 APC 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.

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. Sekine C, et al. 2009. Int Immunol. PubMed
  5. Grujic M, et al. 2010. J. Immunol. 185:1730. PubMed
  6. Hussain MS, et al. 2013. Hum Mol Genet. 22:5199. PubMed
  7. Feng Q, et al. 2014. PLoS One. 9:95927. PubMed
  8. Isobe T, et al. 2014. eLife. 3:1977. PubMed
Product Citations
  1. Patel RP, et al. 2020. Transl Oncol. 14:100917. PubMed
  2. Inclan-Rico JM, et al. 2020. Nat Immunol. 21:1181. PubMed
  3. Tang Z, et al. 2017. Cell Death Differ. 24:2127. PubMed
  4. Preda MB, et al. 2021. Cell Death Dis. 12:566. PubMed
  5. Chugh RM, et al. 2021. Stem Cell Res Ther. 12:388. PubMed
  6. Isobe T, et al. 2014. Elife. 3:1977. PubMed
  7. Sugiura A, et al. 2022. Immunity. 55:65. PubMed
  8. Zhang H, et al. 2022. Eur J Immunol. 52:978. PubMed
  9. Hao J, et al. 2022. Cell Rep. 41:111804. PubMed
  10. Cappellesso F, et al. 2022. Nat Cancer. 3:1464. PubMed
  11. Liu X, et al. 2023. Nat Commun. 14:238. PubMed
  12. Di Benedetto C, et al. 2023. Cancers (Basel). 15: . PubMed
  13. Daian LM, et al. 2023. Int J Mol Sci. 24: . PubMed
  14. Hochrein SM, et al. 2022. Cell Metab. 34:516. PubMed
  15. Roy S, et al. 2022. Endocrinology. 163:. PubMed
  16. Kalus P, et al. 2022. Int J Mol Sci. 23:. PubMed
  17. McCartin C, et al. 2022. Chem Biol Interact. 367:110167. PubMed
  18. Hagege A, et al. 2022. Cell Rep Med. 3:100659. PubMed
  19. Christian DA, et al. 2022. Sci Immunol. 7:eabq7432. PubMed
  20. Shiroshita K, et al. 2022. Cell Rep Methods. 2:100354. PubMed
  21. Giampaolo S, et al. 2023. iScience. 26:106234. PubMed
  22. Battistello E, et al. 2023. Mol Cell. 83:1216. PubMed
  23. Riera-Domingo C, et al. 2023. Sci Adv. 9:eadd5028. PubMed
  24. Perez GI, et al. 2023. Vaccines (Basel). 11:. PubMed
  25. Chatterjee S, et al. 2023. Sci Adv. 9:eade4800. PubMed
  26. Onyshchenko K, et al. 2023. Nat Commun. 14:2087. PubMed
  27. Pang JM, et al. 2023. PLoS One. 18:e0284744. PubMed
  28. Van Bael J, et al. 2023. Int J Mol Sci. 24:. PubMed
  29. Xiao C, et al. 2023. Nat Aging. 3:418. PubMed
  30. Sun K, et al. 2023. Cancers (Basel). 15:. PubMed
  31. Wyrobnik I, et al. 2023. Oncoimmunology. 12:2219164. PubMed
  32. Jiang D, et al. 2023. Nat Commun. 14:3642. PubMed
  33. McCartin C, et al. 2022. Cancers (Basel). 14:. PubMed
  34. Wu N, et al. 2020. Cell Rep. 30:1129. PubMed
  35. Wiley CD, et al. 2019. Cell Rep. 28:3329. PubMed
  36. Schick M, et al. 2022. Nat Commun. 13:281. PubMed
  37. Kuljanin M, et al. 2018. Cell Rep. 25:2524. PubMed
  38. Wu SJ, et al. 2017. Cell Rep. 2.628472222. PubMed
  39. Xu B, et al. 2017. Oncogenesis. 6:e295. PubMed
  40. Donado CA, et al. 2020. Cell Reports. 31(1):107466. PubMed
  41. Cannons JL, et al. 2021. Cell Rep. 37:109804. PubMed
  42. Dai X, et al. 2017. J Cell Mol Med. 10.1111/jcmm.13296. PubMed
  43. Zhang S, et al. 2019. Front Pediatr. 0.485416667. PubMed
  44. Vig S, et al. 2019. Cell Death Dis. 0.63125. PubMed
  45. Xiao C, et al. 2022. iScience. 25:103934. PubMed
  46. McNeal AS, et al. 2021. Elife. 10:. PubMed
  47. Zeriouh W, et al. 2017. PLoS One. 12(2):e0170823. PubMed
  48. Weiser J, et al. 2015. J Infect Dis. 212: 1677 - 1682. PubMed
  49. Cao J, et al. 2019. Theranostics. 0.938194444. PubMed
  50. Moose DL, et al. 2020. Cell Rep. 30:3864. PubMed
  51. Dregalla RC, et al. 2021. Stem Cell Res Ther. 12:547. PubMed
  52. Linnerbauer M, et al. 2022. Front Immunol. 12:800128. PubMed
  53. Wohlhieter CA, et al. 2020. Cell Rep. 33:108444. PubMed
  54. Pease NA, et al. 2021. Cell Reports. 34(12):108888. PubMed
  55. Figueroa-Valdés AI, et al. 2021. Front Bioeng Biotechnol. 9:619930. PubMed
  56. Fujino T, et al. 2021. Nat Commun. 12:1826. PubMed
  57. Patel RP, et al. 2021. Invest Ophthalmol Vis Sci. 62:16. PubMed
  58. Wang J, et al. 2016. Nat Cell Biol. 18: 480-490. PubMed
  59. Sumitomo Y, et al. 2016. Blood. 128: 1614 - 1624. PubMed
  60. Li L, et al. 2016. Cell Death Dis. 7:e2439. PubMed
  61. Zeng H, et al. 2018. Cancer Cell. 34:56:00. PubMed
  62. Lei W, et al. 2020. Sci Rep. 10:7004. PubMed
  63. Deng J, et al. 2021. Cell Death Dis. 12:978. PubMed
  64. Ning X, et al. 2019. Mol Cell. 74:19. PubMed
  65. Vongpipatana T, et al. 2020. J Immunol. 2156:204. PubMed
  66. O'Neill K, et al. 2016. Genes Dev. 30: 973-988. PubMed
  67. Hussain M, et al. 2013. Hum Mol Genet. 22:5199. PubMed
  68. Sheng Y, et al. 2019. Cell Prolif. 52:e12611. PubMed
  69. Tang Z, et al. 2019. Cell Rep. 28:1744. PubMed
  70. Liu L, et al. 2019. Cell Prolif. :e12718. PubMed
  71. Fleischmann M, et al. 2021. Cells. 10:. PubMed
  72. Rausch M, et al. 2021. Cancers (Basel). 13:. PubMed
  73. Lee M, et al. 2022. Nat Commun. 13:1157. PubMed
  74. Laughney AM, et al. 2020. Nat Med. 26:259. PubMed
  75. Hagan AS, et al. 2020. Development. 147:00:00. PubMed
  76. Hickman O, et al. 2016. Br J Cancer. 10.1038/bjc.2016.91. PubMed
  77. Lee Y, et al. 2022. Sci Adv. 8:eabm7688. PubMed
  78. Qiu C, et al. 2022. Front Immunol. 12:764949. PubMed
  79. Saint Fleur–Lominy S, et al. 2018. Cell Rep. 24:3045. PubMed
  80. Kanda K, et al. 2018. JCI Insight. 3:e91316. PubMed
  81. Zhou X, et al. 2019. Cell Rep. 27:1176. PubMed
  82. Aslam MA, et al. 2021. EMBO Rep. 22:e51184. PubMed
  83. Høgh RI, et al. 2020. J Immunol. 1746:204. PubMed
  84. Zhang S, et al. 2020. Front Immunol. 1.757638889. PubMed
  85. Celus W, et al. 2021. Cancer Immunol Res. Online ahead of print. PubMed
  86. Brasacchio D, et al. 2018. Cell Death Dis. 0.681944444. PubMed
  87. Liu L, et al. 2020. Cancer Res. 80:2564. PubMed
  88. Cao W, et al. 2016. Nat Commun. 7:11687. PubMed
  89. Perny M, et al. 2016. Cell Death Dis. 7:e2447. PubMed
  90. Warmuth S, et al. 2022. Oncoimmunology. 10:2004661. PubMed
  91. Jung E, et al. 2017. J Neurosci. 37:6837. PubMed
  92. Cai J, et al. 2021. eLife. 10:00. PubMed
  93. Cheng CC, et al. 2020. Elife. 9:00. PubMed
  94. Feng Q, et al. 2014. PLoS One. 9:95927. PubMed
  95. Ren Y, et al. 2022. J Immunother Cancer. 10:. PubMed
  96. Hills LB, et al. 2021. J Immunol. 206:89. PubMed
  97. Dou R, et al. 2021. Cell Death Dis. 12:206. PubMed
  98. Burclaff J, et al. 2022. Cell Mol Gastroenterol Hepatol. 13:1554. PubMed
  99. Pedicona F, et al. 2022. Sci Signal. 15:eabl7989. PubMed
  100. Li R, et al. 2022. Exp Hematol Oncol. 11:77. PubMed
  101. He C, et al. 2022. BMC Cancer. 22:1100. PubMed
  102. Qiu Z, et al. 2019. Theranostics. 3.650694444. PubMed
  103. Lasrado N, et al. 2022. iScience. 25:103865. PubMed
  104. Forsberg MH, et al. 2021. Stem Cell Res Ther. 12:459. PubMed
  105. Reyes–Uribe P, et al. 2018. Oncogene. 37:4058. PubMed
  106. Lima S, et al. 2018. Autophagy. 1.2375. PubMed
  107. Oh J, et al. 2017. PLoS Biol. 15:e2003352. PubMed
  108. Okita R, et al. 2021. Thorac Cancer. 12:775. PubMed
  109. Li C, et al. 2021. Cell Metabolism. 33(8):1610-1623.e5. PubMed
  110. Kostadinova E, et al. 2016. Sci Rep. 6:30943. PubMed
  111. Sahoo SS, et al. 2021. Nat Med. 27:1806. PubMed
  112. Sun J et al. 2018. Cell stem cell. 23(3):355-369 . PubMed
  113. Sharma P, et al. 2018. Data Brief. 1.975694444. PubMed
  114. Li B, et al. 2019. Oncol Rep. 41:608. PubMed
  115. Muthalagu N, et al. 2020. Cancer Discov. 1.022222222. PubMed
  116. Pothlichet J, et al. 2020. J Clin Invest. 130:2872. PubMed
  117. Wei Z, et al. 2021. Nat Commun. 0.805555556. PubMed
  118. Millington-Burgess SL, et al. 2021. Sci Rep. 11:17678. PubMed
  119. Fernández D, et al. 2016. PLoS One. 11: 0157889. PubMed
  120. Burgener SS, et al. 2019. Cell Rep. 27:3646. PubMed
  121. Hasselluhn MC, et al. 2019. Cell Death Dis. 0.726388889. PubMed
  122. Xu Y, et al. 2021. IUBMB Life. 73:1432. PubMed
  123. Zhang S, et al. 2018. Sci Rep. 8:17066. PubMed
  124. Shinde R, et al. 2018. Nat Immunol. 1.188194444. PubMed
  125. Ho JY, et al. 2021. Mol Ther Methods Clin Dev. 21:237. PubMed
  126. Hirata SI, et al. 2020. Allergy. 75:1939. PubMed
  127. Vickman R, et al. 2016. Mol Cancer Res. 14: 776 - 786. PubMed
  128. Cabet E, et al. 2015. PLoS One. 10: 0137009. PubMed
  129. Rodriguez-Perdigon M, et al. 2022. Macromol Biosci. 22:e2200168. PubMed
  130. Barve A, et al. 2019. Cells. 1.251388889. PubMed
  131. Alamillo E, et al. 2017. Fish Shellfish Immunol. 10.1016/j.fsi.2017.08.036. PubMed
  132. Voss OH, et al. 2018. Curr Protoc Immunol. 120:14.44.1. PubMed
  133. Dower CM, et al. 2018. Mol Cancer Ther. 2.350694444. PubMed
  134. Dong G, et al. 2020. Haematologica. . PubMed
  135. Tulasi DY, et al. 2021. Cell Mol Gastroenterol Hepatol. 11:1437. PubMed
  136. Yamamoto H, et al. 2016. Sci Rep. 6:19204. PubMed
  137. Grossman L, et al. 2017. mSphere. 2:e00305. PubMed
  138. Habtetsion T et al. 2018. Cell metabolism. 28(2):228-242 . PubMed
  139. Woolf N, et al. 2017. Oncogenesis. 10.1038/oncsis.2017.77. PubMed
  140. Parsa S, et al. 2021. Nat Cancer. 1:653. PubMed
  141. Li S, et al. 2021. Front Pharmacol. 12:634115. PubMed
  142. Satcher R, et al. 2015. J Cell Sci. 128: 4629 - 4641. PubMed
  143. Cordo' V, et al. 2022. Nat Commun. 13:1048. PubMed
  144. Mykhailova O, et al. 2021. Transfusion. 61:1247. PubMed
  145. Walle T, et al. 2022. Sci Adv. 8:eabh4050. PubMed
  146. Festag J, et al. 2020. Mol Ther Nucleic Acids. 1.330555556. PubMed
  147. Wallace JG, et al. 2020. Clin Immunol. 210:108311. PubMed
  148. Höhne K, et al. 2016. Open Bio. 6: 160046. PubMed
  149. Yang X, et al. 2020. Immunity. 51(6):983-996.e6.. PubMed
  150. Chen Y, et al. 2020. Front Oncol. 10:767. PubMed
  151. Lu Y, et al. 2018. Cancer Cell. 33:1048. PubMed
  152. Loo Yau H, et al. 2021. Molecular Cell. 81(7):1469-1483.e8. PubMed
  153. Viviane Ponath, Bernd Kaina 2017. PLoS One. 10.1371/journal.pone.0170347. PubMed
  154. Schäfer AL, et al. 2021. Front Immunol. 12:696810. PubMed
  155. Anderson CJ, et al. 2021. Nature. 596:262. PubMed
  156. Agarwal P, et al. 2021. Cell Reports. 36(2):109386. PubMed
  157. Song S, et al. 2021. Mar Drugs. 19: . PubMed
  158. Trigg RM, et al. 2019. Nat Commun. 4.186111111. PubMed
  159. Young MM, et al. 2019. Cell Death Dis. 1.004861111. PubMed
  160. Amend A, et al. 2021. Int J Mol Sci. 22:. PubMed
  161. Chen Y, et al. 2022. Cell Death Dis. 13:822. PubMed
RRID
not an antibody (BioLegend Cat. No. 640919)
AB_2561515 (BioLegend Cat. No. 640920)
AB_2616657 (BioLegend Cat. No. 640941)

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: 8    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