BD, 554420, BD Pharmingen™ APC Rat Anti-Mouse TNF

Reactivity: Mouse (QC Testing)
Isotype: Rat IgG1
Immunogen: Recombinant Mouse TNF
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_398553
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 to APC under optimum conditions, and unconjugated antibody and free APC 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 Cytometry: The APC-conjugated MP6-XT22 antibody can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate TNF-producing cells within mixed cell populations (see figure). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MP6-XT22 antibody with ligand (e.g., recombinant mouse TNF; Cat. No. 554589) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled MP6-XT22 antibody (Cat. No. 554416) prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse cells is APC-R3-34 (Cat. No. 554686). Isotype controls should be used at a comparable concentration to the antibody of interest. ELISA Capture: The purified MP6-XT22 antibody (Cat. No. 554416) is useful as a capture antibody for a sandwich ELISA for measuring mouse TNF protein levels. WB: The MP6-XT22 antibody has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.