Description
Two-color Fluorescent Ig Isotype Cocktail
Containing the following:
Human CD4 PerCP-Cy5.5 Clone:SK3
Mouse mIgG1, ê FITCClone:MOPC-21
Mouse mIgG1, ê PEClone:MOPC-21
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) 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/Th17 Phenotyping Kit Cocktail (Component 51-9006617 of Cat. No. 560752). This phenotyping cocktail is comprised of PerCP-Cy5.5 anti-Human CD4, FITC-Mouse IgG1, κ anti-Human IFN-γ and PE-Mouse IgG1, κ anti-Human IL-17 antibodies. The BD Human Th1/Th17 Phenotyping Kit provides an easy-to-use three-color cocktail of fluorescent antibodies specific for human CD4, IFN-γ (for Th1) 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.
Suggested Companion Products
Stain Buffer (FBS) RUO 500mLCat No: 554656
PerCP-Cy™5.5 Mouse Anti-Human CD4 SK3 (also known as Leu3a) 50Cat No: 341654
HumanTh1/Th17 Phenotyping Kit RUO 50TestsCat No: 560752
Protein Transport Inhibitor (Containing Monensin) RUO 0.7mLCat No: 554724
Fixation Buffer RUO 100mLCat No: 554655
Perm/Wash Buffer RUO 100mLCat No: 554723
BD Cytofix/Cytoperm™ Fixation/Permeablization Kit RUO 250TestsCat No: 554714
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Preparation and Storage
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 with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.The antibody was conjugated with FITC under optimum conditions, and unreacted FITC was removed.Store undiluted at 4°C and protected from prolonged exposure to light.Do not freeze.
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).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 with other multicolor fluorescent antibody cocktails that contain PerCP-Cy5.5 anti-Human CD4 mixed with FITC- and PE-conjugated Mouse 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 Cytofix and Perm/Wash Buffers.
See the protocol for stimulating, fixing, permeabilizing, and staining cells provided in the Technical Data Sheet for the Human Th1/Th17 Phenotyping Kit Cocktail (Component 51-9006617 of Cat. No. 560752).
