BD, 554701, BD Pharmingen™ PE Mouse Anti-Human IFN-γ

Alternative Name: IFNG; Interferon-gamma; Interferon-γ; Type II interferon; MAF
Reactivity: Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Isotype: Mouse IgG1, κ
Immunogen: Human IFN-γ Recombinant Protein
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_395518
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 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 B27 antibody is useful for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IFN-γ producing cells within mixed cell populations (see image). For optimal immunofluorescent staining for flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). For specific methodology, please visit our web site http://www.bdbiosciences.com/us/s/resources, under protocols in "Intracellular Flow". A useful control for demonstrating specificity of staining is the following: pre-block the paraformaldehyde-fixed/saponin-permeabilized cells with unlabeled B27 antibody (Cat. No. 554669/550011) prior to staining. The intracellular cytokine 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 PE-MOPC-21 (Cat. No. 554680); use at comparable concentrations to antibody of interest (e.g., ≤ 0.5 µg mAb/1 million cells). Neutralization: The NA/LE™ B27 antibody (Cat. No. 554698) is useful for neutralization of human IFN-γ bioactivity. A suitable NA/LE™ mouse IgG1 isotype control to match the NA/LE B27 antibody is the 107.3 antibody, (Cat. No. 554721). IP/WB: The PE-conjugated B27 antibody has been reported to be useful for immunoprecipitation studies. The B27 antibody has been reported not to bind to denatured IFN-γ.
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. An isotype control should be used at the same concentration as the antibody of interest. 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. Cy is a trademark of GE Healthcare. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.