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FITC Annexin V
FITC Annexin V
640905 25 tests $70.00 Qty:
    
640906 100 tests $165.00 Qty:
    

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 FITC under optimal conditions. The solution is free of unconjugated FITC.
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 staining of apoptotic cells and analyzing by flow cytometry. The suggested use of this reagent is 5 µl per 1X105 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:
Blue Laser (488 nm)
COA:
Enter Lot#:   
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 FITC 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:

  Publication Library
FITC Annexin V  Image 1
Human T leukemia cell line, Jurkat, non-treated (top) or treated (bottom) with BioLegend’s anti-human CD95 (EOS9.1) mAb (cat. 305704) for 6 hours, then stained with Annexin V-FITC and Propidium Iodide (PI) (cat. 421301)

FITC Annexin V  Image 2


Compare all formats

See FITC spectral data







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. So, we recommend using our 7-AAD Viability Staining Solution (cat. no. 420403/420404) or Propidium Iodide Solution (cat. no. 421301). 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
GeneID: 308
Latest Publications: View the latest Annexin V articles on HighwirePress.com
UniProt: View information about Annexin V on UniProt.org
Keywords: FITC Annexin V, FITC, Annexin A5 , All mammalian species, Flow Cytometry, Biofunctional Assay, Immunology, Antibodies


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Product FAQs

 
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.

http://www.ufpi.br/subsiteFiles/ppgcf/arquivos/files/10.pdf

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.

Can I freeze Annexin V conjugates?

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

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.

For a complete list of FAQs on all our products, visit our FAQ page.
 
Related Products

DescriptionCloneApplications
Annexin V Binding BufferFC, IF
Propidium Iodide SolutionFC
7-AAD Viability Staining SolutionFC


Related Pathways

Pathway
Mechanisims of Cell Death


Related Categories

Category
Cell Biology > Antigens A-Z > Annexin V
Cell Biology > Apoptosis/Tumor Suppressors > Annexin V

*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). 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.

Version: 3 Revision Date: 2013-07-05
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Compare Data Across All Formats
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.

  • Pacific Blue™ Annexin V Apoptosis Detection Kit with PI
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, treated (top) or non-treated (bottom) with LEAF™ purified anti-human CD95 (clone EOS9.1) for 4 hours, then stained with Pacific Blue™ Annexin V Apoptosis Detection Kit with PI.





  • Pacific Blue™ Annexin V
    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





  • FITC Annexin V
    Human T leukemia cell line,

    Human T leukemia cell line, Jurkat, non-treated (top) or treated (bottom) with BioLegend’s anti-human CD95 (EOS9.1) mAb (cat. 305704) for 6 hours, then stained with Annexin V-FITC and Propidium Iodide (PI) (cat. 421301)





  • Alexa Fluor® 647 Annexin V
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, was stimulated (4 hours) with (filled histogram) or without (open histogram) LEAF™ purified anti-CD95 (clone EOS9.1), then stained with Annexin V Alexa Fluor® 647.

  • APC Annexin V
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, was treated (4 hours) with (filled histogram) or without (open histogram) LEAF™ purified anti-CD95 (clone EOS9.1), then stained with Annexin V APC.

  • PE Annexin V
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, was treated (4 hours) with (filled histogram) or without (open histogram) LEAF™ purified anti-CD95 (clone EOS9.1), then stained with Annexin V PE.

  • FITC Annexin V Apoptosis Detection Kit with PI
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, treated (top) or non-treated (bottom) with BioLegend's LEAF™ purified anti-CD95 (clone EOS9.1) mAb (Cat. No. 305704) for 4 hours, then stained with FITC Annexin V Apoptosis Detection Kit with PI (Cat. No. 640914)





  • FITC Annexin V Apoptosis Detection Kit with 7-AAD
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, treated (top) or non-treated (bottom) with BioLegend's LEAF™ purified anti-human CD95 (clone EOS9.1) mAb (Cat. No. 305704) for 4 hours, then stained with FITC Annexin V Apoptosis Detection Kit with 7-AAD (Cat. No. 640922).





  • Purified Annexin V
    Human T cell leukemia cell

    Human T cell leukemia cell line, Jurkat, was treated (4 hours) with (top 2 panels) or without (bottom two panels) LEAF™ purified anti-CD95 (clone EOS9.1), then stained with Annexin V FITC (panels 1 & 3) or with purified recombinant Annexin V to block Annexin V FITC staining (panels 2 & 4).





  • Brilliant Violet 421™ Annexin V
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, was stimulated (4 hours) with (filled histogram) or without (open histogram) LEAF™ purified anti-CD95 (clone EOS9.1), then stained with Annexin V Brilliant Violet 421™.

  • Pacific Blue™ Annexin V Apoptosis Detection Kit with 7-AAD
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, treated (top) or non-treated (bottom) with LEAF™ purified anti-human CD95 (clone EOS9.1) for 4 hours, then stained with Pacific Blue™ Annexin V Apoptosis Detection Kit with 7-AAD.





  • APC Annexin V Apoptosis Detection Kit with 7-AAD
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, treated (top) or non-treated (bottom) with LEAF™ purified anti-human CD95 (clone EOS9.1) for 4 hours, then stained with APC Annexin V Apoptosis Detection Kit with 7-AAD.





  • PE Annexin V Apoptosis Detection Kit with 7-AAD
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, treated (top) or non-treated (bottom) with LEAF™ purified anti-human CD95 (clone EOS9.1) for 4 hours, then stained with PE Annexin V Apoptosis Detection Kit with 7-AAD.





  • Biotin Annexin V
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, was treated with (filled histogram) or without (open histogram) LEAF™ purified anti-CD95 (clone EOS9.1) for 4 hours, then stained with biotinylated Annexin V, followed by Sav-PE.

  • APC Annexin V Apoptosis Detection Kit with PI
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat was treated with (top) or without (bottom) LEAF™ purified anti-human CD95 (clone EOS9.1) for 4 hours, then stained with APC Annexin V Apoptosis Detection Kit with PI.





  • PerCP/Cy5.5 Annexin V
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, was treated with (filled histogram) or without (open histogram) LEAF™ purified anti-CD95 (clone EOS9.1) for 4 hours, then stained with Annexin V PerCP/Cy5.5.

  • Brilliant Violet 510™ Annexin V
    Human T-cell leukemia cell line,

    Human T-cell leukemia cell line, Jurkat, was treated with (filled histogram) or without (open histogram) LEAF™ purified anti-CD95 (clone EOS9.1) for 4 hours, then stained with Annexin V Brilliant Violet 510™.

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