Alexa Fluor® 488-labelled recombinant mouse annexin shows bright green fluorescence (Ex(max): 495nm; Em(max): 519nm).
|Source:||Produced in E. coli. |
|Gene/Protein Identifier:||A000281 (UCSD Signaling Gateway ID)|
|Formulation:||Liquid. Contains 0.02% sodium azide.|
|Specificity:||Binds phosphatidylserine (PS) residues.|
|Application Notes:||Detection of apoptotic cells by flow cytometry.|
|Shipping:||Shipped on Blue Ice|
|Long Term Storage:||+4°C|
|Handling:||Protect from light. Do not freeze.|
|Protocol:||General Protocol - Detection of apoptotic cells by flow cytometry |
- Harvest 2x105 cells per staining and wash cells twice in cold phosphate- buffered-saline (PBS).
- Resuspend cells in 100µl binding buffer (10mM HEPES, pH 7.4, 140mM sodium chloride, 2.5mM calcium dichloride), add 5µl annexin V-Alexa Fluor® 488 to the cells and mix gently.
For detection of necrosis add additionally appropriate amounts of propidium iodide (final concentration 1µg/ml or 1.5µM).
- Incubate for 15 min. at room temperature in the dark.
- Add 400µl binding buffer and analyze the staining in flow cytometry as soon as possible (within an hour).
Figures: Immature mouse B-cells were treated with Fab2-antibodies for 24 hours. Stimulated ((B), after anti-IgM cross-linking) and unstimulated premature mouse B-cells ((A) WEHI231) were stained with Annexin-AlexaFluor® 488 as described  and analyzed in flow cytometry.
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Product Literature References
Engagement of the antigen-receptor on immature murine B lymphocytes results in death by apoptosis
: A. Norvell, et al.; J. Immunol. 154
, 4404 (1995), Abstract