BD, 559502, BD Pharmingen™ PE Rat Anti-Mouse IL-17A

Reactivity: Mouse (QC Testing)
Isotype: Rat LEW, also known as Lewis IgG1, κ
Immunogen: Recombinant Mouse IL-17A Protein
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_397256
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 Recommended Assay Procedure: Immunofluorescent Staining and Flow Cytometric Analysis: The TC11-18H10 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-17A producing cells within mixed cell populations. The PE-conjugated TC11-18H10 antibody (Cat. No. 559502) is especially suitable for these experiments (see image). For optimal immunofluorescent staining and flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.25 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 the specificity of staining is to pre-block paraformaldehyde-fixed/saponin-permeabilized target cells with unlabeled TC11-18H10 antibody (Cat. No. 559501) prior to staining. The intracellular 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 is PE-R3-34 (Cat. No. 554685); use at comparable concentrations to antibody of interest (e.g., ≤ 0.25 µg mAb/1 million cells). OTHER APPLICATIONS 1. ELISA Capture: The purified TC11-18H10 antibody (Cat. No. 555068) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-17A protein levels. Purified TC11-18H10 antibody can be paired with the biotinylated TC11-8H4 (Cat. No.555067) antibody as the detecting antibody, with recombinant mouse IL-17A protein as the standard. The purified TC11-18H10 antibody should be titrated from 0.5 µg/ml to 2.0 µg/ml to determine its optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of mouse IL-17A ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook. 2. Western blotting: The TC11-1810 antibody has been found useful for Western Blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen. 3. Neutralization: TCII-18H10 has been shown to be a neutralizing antibody. Please note that this application is not routinely tested at BD Biosciences.