Flow cytometric analysis of live mouse bone marrow cells using Ly-6G/Ly-6C (Gr-1) (RB6-8C5) Rat mAb (PE-Cy7® Conjugate) (solid line) compared to concentration-matched Rat (LTF-2) mAb IgG2b Isotype Control (PE-Cy7® Conjugate)
#43426 (dashed line).
This Cell Signaling Technology antibody is conjugated to PE-Cy7® and tested in-house for direct flow cytometric analysis in mouse cells.
For optimal flow cytometry results, we recommend 0.25 μg of antibody per test.
Supplied in 10 mM NaH2PO4, 150 mM NaCl, 0.09% NaN3, 0.1% gelatin, pH 7.2. This product is stable for 6 months when stored at 4ºC. 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
Ly-6G/Ly-6C (Gr-1) (RB6-8C5) Rat mAb (PE-Cy7® Conjugate) recognizes endogenous levels of total Ly-6G/Ly-6C (Gr-1) 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.
The Ly-6 complex is a series of genes found on chromosome 15. These genes code for a number of different proteins that can be used as surface markers. The family members vary in their biologic expression and have been shown to be involved in cell signaling and cell adhesion (1). The structure of these proteins includes a motif known as the LU domain that has three loops comprised of disulfide bonds. These bonds are formed by 8 to 10 cysteines that can cause differences in the length of the loops as well as the sequences at each tip (2,4). There are 11 known Ly-6 genes on murine chromosome 15 that code for different proteins. These family members, excluding secreted Ly6/Plaur domain containing 1 coded by the Slurp1 gene, are attached to the cell surface by a GPI anchor near the C terminus. The structure of these proteins may play a role in transmembrane interactions, and downstream signaling cascades (1,2).
Ly-6 proteins have been widely used as differentiation markers on hematopoietic cells. The ability to isolate and express specific Ly-6 antibodies through hybridoma technology has allowed researchers to identify unique proteins (1). These proteins are expressed on subsets of immune cells at different stages of development, such as T cells, B cells, monocytes, granulocytes, and macrophages (1-5).
The 1A8 clone is specific to Ly-6G and is widely used to identify mature granulocytes (3). The RB6-8C5 clone recognized both Ly-6G and Ly-6C, also known as Gr-1, and has been found to express on neutrophils, monocytes, dendritic cells, and T cells (2,3).
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