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.
Product Notices: Product Notices Please refer to www.bdbiosciences.com/us/s/resources for technical protocols. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf. Since applications vary, each investigator should titrate the reagent to obtain optimal results.