BD, 557643, BD Pharmingen™ PE-Cy™7 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_396760
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 PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining and Flow Cytometric Analysis: The PE-Cy7-conjugated B27 antibody (Cat. No. 557643) is useful for multicolor immunofluorescent staining and flow cytometric analysis to identify and enumerate IFN-γ producing cells within mixed cell populations. 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. For specific methodology, please visit the protocols section under "Intracellular Flow" on our web site, http://www.bdbiosciences.com/us/s/resources. Neutralization: The NA/LE B27 antibody (Cat. No. 554698) is useful for neutralization of human IFN-γ bioactivity. IP/WB: The 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 these applications are not routinely tested at BD Biosciences Pharmingen.
Product Notices: Product Notices An isotype control should be used at the same concentration as the antibody of interest. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test). Source of all serum proteins is from USDA inspected abattoirs located in the United States. 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. APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate. APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036). Cy is a trademark of GE Healthcare. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.