BD, 557719, BD Pharmingen™ Alexa Fluor® 488 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_396828
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 Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures: Recommended Assay Procedures Recommended Assay Procedure: The Alexa Fluor® 488-conjugated MP6-XT22 antibody (Cat No. 557719) is available for multicolor immunofluorescent staining and flow cytometric analysis 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 the protocols section or the chapter on intracellular staining for flow cytometric analysis in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MP6-XT22 antibody with a molar excess of ligand (e.g., recombinant mouse TNF; Cat No. 554589) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabelled MP6-XT22 antibody (Cat. No. 554417) prior to staining. The staining technique and 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 and human cells is Alexa Fluor® 488 conjugated R3-34 (Cat. No. 557720); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/ 1 million cells).