Red blood cell lysed peripheral blood mononuclear cells were left unstimulated (left column) or stimulated (right column) for 5 hours with phorbol myristate acetate (PMA) and ionomycin in the presence of brefeldin A. Cells were then fixed and permeabilized using the FIX and PERM® Cell Permeabilization Kit (Cat. no. GAS003 or GAS004) and stained intracellularly with anti-human CD4 APC (Cat. no. MHCD0405) and either anti-human interferon gamma (IFN-gamma) FITC (Cat. no. MHCIFG01) (A) or anti-human tumor necrosis factor alpha (TNF-a) FITC (Cat. no. A18469) (B). Samples were collected using the Attune® Acoustic Focusing Cytometer (blue/red) with 488 nm excitation and 530/30 nm bandpass emission filter to detect FITC. 633 nm excitation and 660/20 nm bandpass emission filter were used to detect APC.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Storage buffer||PBS with 4mg/ml BSA|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Miscellaneous PubMed (MISC)||See 2 publications below|
IFN gamma, also called Type II interferon, macrophage activation factor, and immune interferon is produced primarily by T-lymphocytes and natural killer cells in response to antigens, mitogens, Staphylococcus enterotoxin B, phytohemaglutanin and other cytokines. IFN gamma is a glycoprotein that exists, functionally, as a homodimer of approximately 45 kDa. On SDS-PAGE it appears as a combination of 25, 20 and minor 15.5 kDa bands as a result of differential glycosylation. The biological activity of the IFN gamma homodimer is highly species specific and includes the following: antiviral activity, tumor antiproliferative activity, induction of class I and II MHC, macrophage activation, and enhanced immunoglobulin secretion by B lymphocytes. IFNg is involved in cytokine regulation and also acts synergistically with other cytokines.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Induction of allopeptide-specific human CD4+CD25+ regulatory T cells ex vivo.
MHCIFG01 was used in flow cytometry to investigate antigen specificity of CD4+CD25+ regulatory T cells.
|Jiang S,Camara N,Lombardi G,Lechler RI||Blood (102:2180)||2003|
Human NKT cells mediate antitumor cytotoxicity directly by recognizing target cell CD1d with bound ligand or indirectly by producing IL-2 to activate NK cells.
MHCIFG01 was used in flow cytometry to elucidate the mechanisms of NKT cell cytotoxicity and activation of NK cells using alpha-Galactosylceramide.
|Metelitsa LS,Naidenko OV,Kant A,Wu HW,Loza MJ,Perussia B,Kronenberg M,Seeger RC||Journal of immunology (Baltimore, Md. : 1950) (167:3114)||2001|