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Safety Data Sheet (SDS) Pacific Blue Annexin V     Product Data Sheet (PDF)    
Pacific Blue™ Annexin V
3804585 25 tests $100.00       
3804590 100 tests $245.00       
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

Application Notes:

Annexin V Staining
1. Wash cells twice with cold cell staining buffer and then resuspend cells in Annexin V Binding Buffer 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 or 7-AAD.
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 to each tube. Analyze by flow cytometry.

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.

Application References:

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

Human T-cell leukemia cell line,

Human T-cell leukemia cell line, Jurkat, stimulated (4 hours) with (bottom) or without (top) LEAF™ purified anti-CD95, clone EOS9.1, then stained with Annexin V Pacific Blue™ and viability dye 7-AAD





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 is recommended for use with Annexin V staining.

Annexin V binding alone cannot differentiate between apoptotic cells and necrotic. So, we recommend using our 7-AAD Viability Staining Solution or Propidium Iodide Solution . 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.

Other Names: Annexin A5

This product is provided under an agreement between Life Technologies Corporation and Sony Biotechnology Inc. and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications, and corresponding non-U.S. equivalents, owned by and/or licensed to Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research and/or development conducted by the buyer (whether the buyer is an academic or for profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to (1) use of the product or its components in manufacturing, (2) use of the product or its components for therapeutic, diagnostic or prophylactic purposes, or (3) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Molecular Probes Detection Technologies, Attn: Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.