BD, 559327, 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)
Vol. Per Test: 20 µl
RRID: AB_397224
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 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). This 100 Test Size formulation of the PE-conjugated B27 antibody has been pre-titrated to assure effective intracellular detection of human IFN-γ using 20 µl per 1 x 10e6 cells. For specific methodology, please visit our website, http://www.bdbiosciences.com/us/s/resources and refer to the protocols section under "Cytokines (Intracellular Staining)" or "Intracellular Flow". A suitable mouse IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponinpermeabilized human cells is also available in a 100 Test Size formulation PE-MOPC-21 (Cat. No. 559320). 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. 554699/550011) prior to staining. The intracellular cytokine staining technique and blocking controls are described in detail by C. Prussin and D. Metcalfe. 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. 2. Incubate the cell suspension for 15 minutes (4°C, in the dark). 3. Wash twice in 100 µl of 1X Perm/Wash™ Buffer.