BD, 559321, BD Pharmingen™ PE 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)
Vol. Per Test: 20 µl
RRID: AB_397219
Storage Buffer: Aqueous buffered solution containing BSA and ≤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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining and Flow Cytometric Analysis: The PE conjugated MAb11 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify and enumerate TNF producing cells within mixed cell populations (see Figure). This 100 Test Size formulation of the PE-conjugated MAb11 antibody has been pre-titrated to assure effective intracellular detection of human TNF using 20 µl/1 x 10^6 cells. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit the protocols section under "Cytokines (Intracellular Staining)" and "Intracellular Flow" posted on our web site, http://www.bdbiosciences.com/us/s/resources. Important Note: This pre-titered antibody solution does not contain a cell permeabilization agent. It is necessary to include a cell permeabilization agent when using the pre-titered antibody solution to stain fixed and permeabilized cells. Perm/Wash™ Buffer (Cat. No. 554723) contains the permeabilization agent saponin and is useful for this purpose as described in the USAGE section below. USAGE 1. Resuspend 1 x 10^6 fixed and permeabilized cells in 20 µl of the pre-titered antibody solution and 30 µl of 1X Perm/Wash™ Buffer (Cat. No. 554723). 2. Incubate the cell suspension for 15 minutes (at RT or 4°C). 3. Wash twice in 100 µl of 1X Perm/Wash™ Buffer (Cat. No. 554723).
Product Notices: Product Notices This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test). An isotype control should be used at the same concentration as the antibody of interest. Source of all serum proteins is from USDA inspected abattoirs located in the United States. 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.