Flow cytometric analysis of live mouse splenocytes, untreated (left column) or treated with LPS #14011 (500 ng/ml, 3 d; right column), using CD86/B7-2 (GL-1) Rat mAb (APC Conjugate) (top row) or concentration-matched Rat Isotype Control (APC Conjugate) (bottom row), and co-stained with CD19 (1D3) Rat mAb (FITC Conjugate) #53343.
|Source/Isotype||Rat IgG2a kappa|
This Cell Signaling Technology antibody is conjugated to APC and tested in-house for direct flow cytometric analysis in mouse cells.
For optimal flow cytometry results, we recommend 0.06 μg of antibody per test.
Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. Do not aliquot the antibody. Protect from light. Do not freeze.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.
NOTE: Count cells using a hemocytometer or alternative method.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to Immunostaining.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
posted June 2017
revised June 2020
Protocol Id: 1504
CD86/B7-2 (GL-1) Rat mAb (APC Conjugate) recognizes endogenous levels of total CD86/B7-2 protein. This antibody detects an epitope within the extracellular domain.
This monoclonal antibody was purified from tissue culture supernatant via affinity chromatography. The purified antibody was conjugated under optimal conditions, with unreacted dye removed from the preparation.
CD80 (B7-1, BB1) and CD86 (B7-2, B70) are members of the B7 family of cell surface ligands that regulate T cell activation and immune responses. CD80 is expressed on activated antigen presenting cells, including dendritic cells, B cells, monocytes, and macrophages. CD86 is expressed on resting monocytes, dendritic cells, activated B lymphocytes, and can be further upregulated in the presence of inflammation (1-3). CD80 and CD86 are ligands for CD28, which functions as a T cell costimulatory receptor. Interaction of CD28 with CD80 or CD86 provides the second signal required for naïve T cell activation, T cell proliferation, and acquisition of effector functions (3-7). Alternatively, CD80 and CD86 also act as ligands to CTLA-4, which results in the downregulation of T cell activity (3,7-9).
The GL-1 antibody is used as a marker for CD86 expression on B cells, macrophages, and dendritic cells.
Explore pathways + proteins related to this product.