BD, 560802, BD Pharmingen™ Three-color Fluorescent Ig Isotype Cocktail with Human CD4+ PerCP-Cy™5.5

Reactivity: Human (Reported)
Application: Flow cytometry (Tested During Development)
Regulatory Status: RUO
RRID: AB_2034027
Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
Description: Description This multicolor cocktail contains a mixture of PerCP-Cy5.5-anti-Human CD4 antibody (Clone SK3) and FITC-Mouse IgG1, κ (MOPC-21) and PE-Mouse IgG1, κ (MOPC-21) and APC-Rat IgG1, κ (R3-34) isotype controls. This fluorescent Ig isotype control cocktail serves as a negative control for immunofluorescent staining and flow cytometric analysis using the BD Human Th1/Th2/Th17 Phenotyping Kit Cocktail (Component 51-9006615 of Cat. No. 560751). This phenotyping cocktail is comprised of PerCP-Cy5.5 anti-Human CD4, FITC-Mouse IgG1, κ anti-Human IFN-γ, APC-Rat IgG1, κ anti-Human IL-4, and PE-Mouse IgG1, κ anti-Human IL-17 antibodies. The BD Human Th1/Th2/Th17 Phenotyping Kit provides an easy-to-use four-color cocktail of fluorescent antibodies-specific for human CD4, IFN-γ (for Th1), IL-4 (for Th2) and IL-17A (for Th17) that enables researchers to identify and characterize the nature of T helper cell types by multicolor immunofluorescent staining and flow cytometric analysis.
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 PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.
Recommended Assay Procedures: Recommended Assay Procedures The multicolor Ig isotype control cocktail is designed to control for nonspecific staining by fluorescent antibodies directed against cellular antigens found in target CD4+ T cells that have been fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723), or the BD Cytofix/Cytoperm™ Fixation/Permeabilization Solution Kit (Cat. No. 554714).  Nonspecific staining by fluorescent antibodies may be due to nonspecific immunoglobulin and/or fluorochrome- mediated binding to cellular molecules including Fc receptors for immunoglobulin. In addition, this multicolor Ig isotype cocktail can be used for negative control staining for other multicolor fluorescent antibody cocktails that contains PerCP-Cy™5.5 anti-Human CD4 mixed with FITC- and PE-conjugated Mouse IgG1, κ and APC-Rat IgG1, κ antibodies. As such, the cocktail can provide negative controls for the immunofluorescent staining with fluorescent antibodies specific for other intracellular markers (eg, cytokines, chemokines, and/or transcription factors) expressed by CD4+ T cells that are prepared using BD Cytofix™ Fixation buffer and BD Perm/Wash™ Perm/Wash Buffer. See the protocol for stimulating, fixing, permeabilizing, and staining cells provided in the Technical Data Sheet for the Human Th1/Th2/Th17 Phenotyping Kit (Cat. No. 560751). BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.   However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.