BD, 554479, BD Pharmingen™ PE Rat Anti-Mouse IL-12 (p40/p70)

Reactivity: Mouse (QC Testing)
Isotype: Rat IgG1
Immunogen: CHO-expressed recombinant mouse IL-12 p70 protein
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_395420
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 and Flow Cytometric Analysis: The C15.6 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-12 producing cells within mixed cell populations. The PE- and APC-conjugated C15.6 antibodies are especially suitable for these experiments (see figure). For optimal immunofluorescent staining and flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). For specific methodology, see The Immune Function Handbook at our website at www.bdbiosciences.com. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the PE-C15.6 antibody with excess ligand prior to staining, or 2) pre-block paraformaldehyde-fixed/saponin-permeabilized cells with unlabeled C15.6 antibody (e.g., Cat. No. 554477) prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG1 isotype control immunoglobulin for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse or human cells is PE-R3-34 immunoglobulin (Cat. No. 554685); use at comparable concentrations to antibody of interest. ELISA: The purified C15.6 antibody (Cat. No. 551219) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-12 p40 protein levels. The purified C15.6 antibody can be paired with the biotinylated C17.8 antibody (Cat. No. 554476) as the detection antibody, with recombinant mouse IL-12 p40 protein (Cat. No. 554594) as the standard. For testing mouse IL-12 p40 in complex biological fluids such as serum or plasma, our mouse IL-12 specific OptEIA™ sandwich ELISA set is recommended (Cat. No. 555165).