Intracellular Staining by Flow Cytometry
|Detection of CXCL1/GRO alpha /KC/CINC‑1 in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes either (A) treated with 1 μg/mL LPS and Brefeldin A for 4 hours or (B) untreated were stained with Rabbit Anti-Mouse CXCL1/GRO alpha /KC/CINC‑1 Monoclonal Antibody (Catalog # MAB4532) followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110) and Rat Anti-Mouse TNF‑ alpha Fluorescein‑conjugated Monoclonal Antibody (Catalog # IC410F). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012).|
KC, a member of the alpha (CXC) chemokine subfamily, was initially identified as an immediate early gene induced in mouse fibroblasts by platelet‑derived growth factor. KC cDNA encodes a 96 amino acid (aa) residue precursor protein with a predicted secretory signal peptide that is removed to yield the mature protein. The protein sequence of mouse KC shows approximately 63% identity to that of mouse MIP-2. KC is also approximately 60% identical to the human GROs. It has been suggested that mouse KC and MIP-2 are the orthologs of the human GROs and rat CINCs. In addition to mouse fibroblasts, KC is expressed in macrophages and endothelial cells. Mouse KC is a potent neutrophil attractant and activator. The functional receptor for KC has been identified as CXCR2. Based on the pattern of KC expression in a number of inflammatory disease models, KC appears to have an important role in inflammation. KC was found to be involved in monocyte arrest on atherosclerotic endothelium and may also play a pathophysiological role in Alzheimer’s disease. Many chemokines are substrates for selective proteolysis at the amino-terminus by various proteases including dipeptidyl peptidase IV or matrix metalloproteases, resulting in truncated chemokine isoforms with different (both enhanced or reduced) bioactivities. The naturally occurring 68 aa N-terminal truncated isoform of mouse KC is reported to be a more potent synergistic growth stimulant for CFU-GM.
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Flow Cytometry: Mouse CXCL1/GRO alpha /KC/CINC‑1 Antibody [MAB4532]