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Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq. DescriptionThe 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 and CD45R/B220) reveals a subset of total BM cells staining with RAM34 that can express undetectable to low levels of the mentioned lineage markers. Applications ReportedFor research use only, not for diagnostic or therapeutic use. This RAM34 antibody has been reported for use in flow cytometric analysis. Applications TestedThis RAM34 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This can be used at less than or equal to 2 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. For more detailed information on staining with RAM34, please reference the following publication PubMed. References
Osawa M, Hanada K, Hamada H, Nakauchi H. 1996. Long-term lymphohematopoietic reconstitution by a single CD34-low/negative hematopoietic stem cell. Science. 273(5272):242-5. Related ProductsCat. 51-4321 Alexa Fluor® 647 Rat IgG2a Isotype Control Alexa Fluor® and Pacific Blue® are registered trademarks of and licensed under patents assigned to Molecular Probes, Inc. for research use only. This product is subject to an agreement between Molecular Probes, Inc. and eBioscience, and the manufacture, use, sale or import of this product may be subject to one or more U.S. patents, pending applications and corresponding foreign equivalents, owned by Molecular Probes, Inc. (a wholly owned subsidiary of Invitrogen Corp). The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product for life science research or as an ASR. The buyer cannot use this product for manufacturing or for any other screening (specifically including use in combination with microarrays or High Content Screening) or testing purpose, other than as an ASR. For information on purchasing a license to this product for purposes other than life science research or use as an ASR, contact Molecular Probes, Inc. |
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