BD, 559503, BD Pharmingen™ PE Hamster Anti-Rat/Mouse TNF

Reactivity: Mouse, Rat (QC Testing)
Isotype: Armenian Hamster IgG, λ1
Immunogen: Recombinant mouse TNF protein
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_397257
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 The PE-conjugated TN3-19.12 antibody can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate rat 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 TN3-19.12 antibody with a molar excess of ligand prior to staining, or 2) pre-block the fixed/permeabilized cells with unconjugated TN3-19.12 antibody (Cat. No. 557516) prior to staining. The staining technique and blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable hamster IgG1, λ isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse and human cells is PE-G235-2356 (Cat. No. 554711); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/ 1 million cells). OTHER APPLICATIONS In vitro neutralization: The NA/LE™ format of the TN3-19.12 antibody (Cat. No. 557370) is useful for neutralization of mouse TNF bioactivity. IP/WB: The TN3-19.12 antibody has been reported to be useful for immunoprecipitation and Western blot. Please note that this application is not routinely tested at BD Biosciences Pharmingen. ELISA: The purified TN3-19.12 antibody (Cat. No. 557516) can be used as a capture antibody for a sandwich ELISA that measures TNF protein levels.
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. 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.