BD, 554443, BD Pharmingen™ PE Hamster Anti-Mouse/Rat MCP-1

Reactivity: Mouse, Rat (QC Testing)
Isotype: Armenian Hamster IgG1, κ
Immunogen: Heparin-purified CHO-expressed mouse MCP-1
Application: Flow cytometry (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_395394
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining: The 2H5 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate MCP-1 producing cells within mixed cell populations. For optimal immunofluorescent staining for flow cytometric analysis, the anti-cytokine antibody should be titrated (≤  0.5 µg mAb/million cells). For specific methodology, please visit the protocols sections, or the chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our website, www.bdbiosciences.com. A useful control for demonstrating specificity of staining is either of the following: 1) preblock the conjugated 2H5 antibody with a ligand (e.g., recombinant mouse MCP-1; Cat No. 554590 for mouse or recombinant rat MCP-1; Cat. No. 555110 for rat) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled 2H5 antibody (Cat. No. 554441) prior to staining. The staining technique and blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable hamster IgG isotype control for assessing the level of background staining on fixed/permeabilized mouse cells is PE-G235-2356 (Cat. No. 554711); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells). ELISA: Purified 2H5 antibody (Cat. No. 554441) can be used as the capture antibody in a sandwich ELISA for measuring mouse MCP-1 levels in conjunction with the biotinylated 4E2/MCP mouse antibody (Cat. No. 554444) as the detection antibody, and recombinant mouse MCP-1 (Cat. No. 554590) as the standard. Neutralization: The NA/LE™ 2H5 antibody is useful for neutralization of mouse MCP-1 bioactivity.