BD, 554514, BD Pharmingen™ APC Mouse Anti-Human TNF

Alternative Name: Tumor necrosis factor alpha; TNF-a; TNF-α; TNFSF2; Cachectin
Reactivity: Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Isotype: Mouse IgG1, κ
Immunogen: Recombinant Human TNF
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_398566
Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. 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.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining and Flow Cytometry: The APC-conjugated MAb11 antibody can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate TNF-producing cells within mixed cell populations (see image). 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 the protocols section under "Cytokines (Intracellular Staining)" or "Intracellular Flow" at our website, http://www.bdbiosciences.com/us/s/resources. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MAb11 antibody with a ligand (e.g., recombinant human TNF; Cat No. 554618) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled MAb11 antibody (Cat. No. 554510) prior to staining. The staining technique and blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable mouse IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is APC-MOPC-21 (Cat. No. 554681); use at comparable concentrations to antibody of interest.