BD, 557729, BD Pharmingen™ Alexa Fluor® 647 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)
Vol. Per Test: 5 µl
RRID: AB_396837
Storage Buffer: Aqueous buffered solution containing BSA and ≤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® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining and Flow Cytometric Analysis: The FITC-, PE-, APC-, PE-Cy7-, Alexa Fluor® 488-, and Alexa Fluor® 647-conjugated B27 antibodies (Cat. No. 554700, 554701, 554702, 557643, 557718, and 557729) are  useful for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IFN-γ producing cells within mixed cell populations. For optimal immunofluorescent staining this antibody should be used at 5 µl per test. 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. 554699) 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 Alexa Fluor® 647-MOPC-21 (Cat. No. 557732); use at 5 µl/test. 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 107.3 antibody (Cat. No. 554721). IP/WB: The unconjugated B27 antibody has been reported to be useful for immunoprecipitation studies. The B27 antibody has been reported not to bind to denatured IFN-γ. Please note that this application is not routinely tested at BD Biosciences Pharmingen.