Optimized for flow cytometry and fluorescence microscopy applications. A RAPID™ protocol has been developed for Annexin V-FITC binding directly in tissue culture media. This obviates the need for tedious centrifugation and wash steps, that increase the occurrence of mechanical membrane disruption. In addition, since apoptosis is a dynamic process that is ongoing once cells are removed from culture conditions and continues throughout experimental processing, the RAPID™ protocol is recommended for the detection of cells in early apoptosis.
Catalogue Number
PF032
Brand Family
Calbiochem®
Materials Required but Not Delivered
• PBS (137 mM NaCl, 2.7 mM KCl, 4.3 mM Na2HPO4-7H2O, 1.4 mM KH2PO4, adjust pH to 7.4) • 2-20 µl, 20-200 µl, and 200-1000 µl precision pipetters with disposable tips • Microcentrifuge tubes • Adjustable speed microcentrifuge • Glass microscope slides (for microscope analysis only) • Glass coverslips (for microscope analysis only) • dH2O • Ice • Fluorescence microscope or flow cytometer
References
References
Frey, T. 1997. Cytometry28, 253. Darzynkiewicz. Z., et al. 1997. Cytometry27, 1. Boersma, A.W.M., et al. 1996. Cytometry24, 123. Wyllie, A. H. 1993. Br. J. Cancer67, 205. Darzynkiewicz, Z., et al. 1992. Cytometry13, 795. Fadok, V.A., et al. 1992. J. Immunology148, 2207. Kerr, J.F.R., et al. 1972. Cancer26, 239.
Product Information
Detection method
Fluorometric
Declaration
Sold under license of U.S. Patent 5,834,196.
Form
100 Tests
Format
Flow cytometry or fluorescence microscopy
Kit contains
Annexin V-FITC, 5X Binding Buffer, Propidium Iodide, RAPID™ Media Binding Reagent, and a user protocol.
Positive control
Apoptotic Jurkat cells (dexamethasone or anti-Fas treated)
Applications
Key Applications
Flow Cytometry Immunofluorescence
Biological Information
Assay time
15 - 30 min
Sample Type
Intact cells
Species Reactivity
A Broad Range Of Species
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
R Phrase
R: 36
Irritating to eyes.
S Phrase
S: 22-24
Do not breathe dust. Avoid contact with skin.
Product Usage Statements
Intended use
The Calbiochem® Annexin V-FITC Apoptosis Detection Kit is a non-isotopic system for the identification of cell membrane alterations that accompany programmed cell death. Detection may be performed by flow cytometry or by fluorescence microscopy.
Storage and Shipping Information
Ship Code
Blue Ice Only
Toxicity
Multiple Toxicity Values, refer to MSDS
Storage
+2°C to +8°C
Storage Conditions
Annexin V-FITC kit components are shipped on gel ice pack. Upon receipt, store kit at 4°C. To avoid reagent loss in tube caps, briefly pulse spin all small tubes before removing caps. See Precautions and Recommendations.
Do not freeze
Yes
Packaging Information
Transport Information
Supplemental Information
Kit contains
Annexin V-FITC, 5X Binding Buffer, Propidium Iodide, RAPID™ Media Binding Reagent, and a user protocol.
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