Description: The RAM34 monoclonal antibody reacts with mouse CD34, also known as mucosialin. It has been reported that the RAM34 antibody can be used to detect CD34+Sca-1+c-Kit+ cells. Upon further optimization, we have determined that the most effective RAM34 formats for this combination of antigen detection are biotin, FITC, and Alexa Fluor 647. Preliminary data suggest that some antibody conjugates (such as PE and APC fluorochromes) may negatively impact detection of CD34. Interestingly, the PE RAM34 conjugate is suitable for the staining of epithelial cells (NIH3T3).
CD34, a highly glycosylated (approximately 90-120 kDa) member of the sialomucin family, is expressed by capillary endothelial cells, BM stoma and a small subpopulation of mouse bone marrow cells. RAM34 has been used to purify mouse HSCs to near homogeneity. Unlike in humans, primitive adult mouse bone marrow HSCs were detected in the CD34 low to negative fraction.CD34 expressed on endothelial cells is a ligand for CD62L and plays a role in adhesion. Simultaneous staining of mouse bone marrow cells with a cocktail of antibodies to lineage markers (CD3, CD11b, Ly6G, TER-119 andCD45R/B220) reveals a subset of total BM cells staining with RAM34 that can express undetectable to low levels of the mentioned lineage markers.
Note: When staining with the RAM34 antibody an incubation time of 90 minutes is recommended to obtain the optimal signal to noise signal.
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