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. CD34 is a highly glycosylated (approximately 90-120 kDa) member of the sialomucin family and is expressed by capillary endothelial cells, bone marrow stroma, and a small subpopulation of mouse bone marrow cells. RAM34 has been used to purify mouse hematopoietic stem cells (HSC) to near homogeneity. 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 cells that stain with the RAM34 antibody and express undetectable to low levels of the indicated lineage markers. Applications Reported: This RAM34 antibody has been reported for use in flow cytometric analysis. Applications Tested: This RAM34 antibody has been tested by flow cytometric analysis of mouse bone marrow cells. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.Note: When staining with the RAM34 antibody an incubation time of 90 minutes is recommended to obtain the optimal signal to noise ratio. For more detailed information on staining with RAM34, please refer to Ema et al., listed under References. eFluor® 450 is an alternative to Pacific Blue®. eFluor® 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 405 nm; Emission: 445 nm; Laser: Violet Laser. Filtration: 0.2 µm post-manufacturing filtered. CD34 is a highly glycosylated, monomeric, 111-115 kDa surface protein that is present on many stem cell populations. CD34 is a stem cell marker although its expression on human hematopoietic stem cells is reversible. CD34 may serve as a surface receptor that undergoes receptor-mediated endocytosis and regulates adhesion, differentiation, and proliferation of hematopoietic stem cells and other progenitors. CD34 expression is likely to represent a specific state of hematopoietic development that may have altered adhering properties with expanding and differentiating capabilities in both in vitro and in vivo conditions. CD34 is possibly an adhesion molecule with a putative role for mediating the attachment of stem cells to the bone marrow extracellular matrix or directly to stromal cells. Further, CD34 could act as a scaffold for the attachment of lineage specific glycans, allowing stem cells to bind to lectins expressed by stromal cells or other marrow components. CD34 is thought to have a role in presenting carbohydrate ligands to selectins. The intracellular chain of the CD34 antigen is a site of phosphorylation by activated protein kinase C suggesting a putative role in signal transduction. Diseases associated with CD34 dysfunction include dermatofibrosarcoma and neurofibroma.
|4° C, store in dark, DO NOT FREEZE!|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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