Pacific Blue™ Annexin V

Pricing & Availability
Other Names
Annexin A5
Ave. Rating
2 reviews
Product Citations
publications
PB_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 at 37°C, then stained with Annexin V- Pacific Blue™ for 15 minutes at 37°C in Annexin V Binding buffer. Helix NP Green (Cat. No. 425303 at 1.25 nM) was added 5 minutes at room temperature prior to running tubes.
  • PB_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 at 37°C, then stained with Annexin V- Pacific Blue™ for 15 minutes at 37°C in Annexin V Binding buffer. Helix NP Green (Cat. No. 425303 at 1.25 nM) was added 5 minutes at room temperature prior to running tubes.
See Pacific Blue™ spectral data
Cat # Size Price Quantity Avail. Save
640917 25 tests $105
Check Availability


Need larger quantities of this item?
Request Bulk Quote
640918 100 tests $250
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

Product Details

Reactivity
All mammalian species
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and 0.2% (w/v) BSA (origin USA).
Preparation
The purified protein was conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration
Lot-specific (please contact technical support for concentration and total µg amount, or use our Lookup tool if you have a lot number.)
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.

* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome.


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

View full statement regarding label licenses
Excitation Laser
Violet Laser (405 nm)
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 1x10e6 cells/ml.
2. Transfer 100 µl of cell suspension in 5 ml test tube.
3. Add 5 µl of Pacific Blue ™ 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.

Application References

(PubMed link indicates BioLegend citation)
  1. Gordy C, et al. 2011 Blood 117:618. PubMed
  2. Jia W, et al. 2011. J. Immunol. 186:5313. PubMed
  3. Naegele M, et al. 2012. J. Neuroimmunol. 242:60. PubMed
  4. Jenke AC, et al. 2013. PLoS One. 8:e55636. PubMed
  5. Oh J, et al. 2013. J Exp Med. 210:1069. PubMed
  6. Hasan S, et al. 2013. Blood. 122:1464. PubMed
  7. Zhao BB, et al. 2013. PLoS One. 8:77008. PubMed
  8. Mihaly SR, et al. 2014. PLoS One. 9:94982. PubMed
  9. Miyazawa M, et al. 2014. Mol Biol Cell. 25:2116. PubMed
  10. Yabas M, et al. 2014. J Biol Chem. 289:19531. PubMed
  11. Burbulla LF, et al. 2014. Cell Death Dis. 5:1180. PubMed
Product Citations
  1. He W 2011. J Immunol. 186:5313. PubMed
  2. Gordy C, et al. 2011. Blood. 117:618. PubMed
  3. Martel C, et al. 2011. PLoS One. 6:e18812. PubMed
  4. Naegele M, et al. 2012. J Neuroimmunol. 242:60. PubMed
  5. Jenke A, et al. 2013. PLoS One. 8:e55636. PubMed
  6. Oh J, et al. 2013. J Exp Med. 210:1069. PubMed
  7. Hasan S, et al. 2013. Blood. 122:1464. PubMed
  8. Zhao B, et al. 2013. PLoS One. 8:77708. PubMed
  9. Mihaly S, et al. 2014. PLoS One. 9:94982. PubMed
  10. Tsuji M 2014. Mol Biol Cell. 25:2116. PubMed
  11. Yabas M, et al. 2014. J Biol Chem. 289:19531. PubMed
  12. Burbulla L, et al. 2014. Cell Death Dis. 5:1180. PubMed
  13. Lao Z, et al. 2015. PLoS One. 10: 0133895. PubMed
  14. Yabas M, et al. 2016. PLoS One. 11: 0146774. PubMed
  15. Peiris T, et al. 2016. Development. 143: 1697 - 1709.. PubMed
  16. Mascarenhas M, et al. 2016. Blood. 127: 2298 - 2309. PubMed
  17. Khan E, et al. 2016. Sci Rep. 6:38104. PubMed
  18. Guentsch A, et al. 2017. Mol Cell Biol. 37: e00236-16. PubMed
  19. Silginer M, et al. 2017. Cell Death Dis. 10.1038/cddis.2017.171. PubMed
  20. Hejna M, et al. 2017. Sci Rep. 10.1038/s41598-017-12165-1. PubMed
  21. Petrillo C 2018. Cell stem cell. 23:820. PubMed
Publication Library
RRID
AB_1279046 (BioLegend Cat. No. 640917)
AB_1279044 (BioLegend Cat. No. 640918)

Related FAQs

Can I freeze Annexin V conjugates?

It should not be frozen as it will lead to loss of biological activity due to dimerization.

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.

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.

How is your Annexin made and what sequence does it cover?

It is made in E. coli, covering human aa Met1-Asp320.

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.

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.

Go To Top Version: 4    Revision Date: 05/23/2018

For research use only. Not for diagnostic use. Not for resale. BioLegend will not be held responsible for patent infringement or other violations that may occur with the use of our products.

 

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/ordering#license). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products, reverse engineer functionally similar materials, or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.

 

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.

Login/Register
Forgot your password? Reset Password
Request an Account