Confocal immunofluorescent analysis of rat oligodendrocyte progenitor cells using A2B5 Mouse mAb and an anti-mouse IgM secondary antibody (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
It is strongly recommended that these directions be followed for staining of live, unpermeabilized cells when using this antibody. Dilute primary antibody 1:100 in culture medium containing 0.1% sodium azide*. Aspirate cell culture medium, add diluted antibody and incubate cells for 30 minutes at 37ºC. Aspirate antibody solution, rinse gently with pre-warmed PBS, and fix with methanol-free, 4% formaldehyde diluted in PBS for 15 minutes at 37ºC. Rinse cells with PBS 3 times for 5 minutes each. For best results, detect with a fluorochrome-conjugated anti-mouse IgM secondary antibody (per manufacturer's recommendation, diluted in PBS). Prior to imaging, rinse samples in PBS 3 times for 5 minutes each and mount in an anti-fade reagent (ProLong® Gold Antifade Reagent #9071).
* sodium azide is included to prevent capping.
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
A2B5 Mouse mAb recognizes endogenous levels of A2B5 antigen. A2B5 Mouse mAb has been validated for use with the recommended staining protocol. Fixation of cells prior to staining will lead to artifacts. Based on literature publications, this antibody is predicted to cross-react with chicken, rat, bovine, and human samples.Species Reactivity:
Monoclonal antibody is produced by immunizing animals with embryonic chicken retinal cells.
A2B5 Mouse mAb recognizes a cell surface ganglioside epitope that has been utilized as a marker for identification of various cell types. A2B5 Mouse mAb has been used to mark specific cell populations such as neuroendocrine cells, thymic epithelial cells (1), and glial precursors that give rise to type II astrocytes and oligodendrocytes (2-4).
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