Description: The MR1 monoclonal antibody reacts with mouse CD154, a 39 kDa transmembrane glycoprotein also known as CD40 ligand (CD40L). CD154 is expressed transiently by activated T cells. Through its binding to CD40 on antigen presenting cells (APC) including B cells, monocytes/macrophages, and dendritic cells, it serves a crucial function in T cell-APC cognate interaction. CD154-interaction with CD40 transduces signals for T-dependent B cell activation and induces B cells to enter the cell cycle. For staining for flow cytometric analysis, it is important to stimulate enriched T cells or enriched CD4 cells (using depletion strategy) prior to staining with MR1. Applications Reported: The MR1 antibody has been reported for use in flow cytometric analysis. It has also been reported to inhibit binding of CD154 to CD40 and activation of B cells. Applications Tested: This MR1 antibody has been tested by flow cytometric analysis of stimulated mouse splenocytes. This can be used at less than or equal to 0.5 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. Endotoxin: Less than 0.001 ng/ug antibody as determined by the LAL assay.Storage and handling: Use in a sterile environment. Filtration: 0.2 µm post-manufacturing filtered. Purity: Greater than 90%, as determined by SDS-PAGE. Endotoxin Level: Less than 0.001 ng/µg antibody, as determined by LAL assay. Aggregation: Less than 10%, as determined by HPLC. CD40 Ligand (CD40-L), or CD154, is a membrane glycoprotein and differentiation antigen expressed on the surface of T cells. The CD40 Ligand stimulates B cell proliferation and secretion of all immunoglobulin isotypes in the presence of cytokines. CD40 Ligand has been shown to induce cytokine production and tumoricidal activity in peripheral blood monocytes. It also co-stimulates proliferation of activated T cells and this is accompanied by the production of IFN-gamma, TNF-alpha, and IL-2.
|CD154 (CD40 Ligand)|
|Flow Cytometry, Functional Assay, Neutralization|
|PBS with no preservative; pH 7.2|
For Research Use Only.
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