C57BL/6 splenocytes were left unstimulated or stimulated for 5 hours with phorbol myristate acetate (PMA) and ionomycin in the presence of brefeldin A. Cells were then surface-stained with anti-mouse CD4 FITC (Cat. No. MCD0401) and subsequently fixed and permeabilized using the FIX and PERM® Cell Permeabilization Kit (Cat. nos. GAS003 or GAS004) and stained intracellularly with anti-mouse interferon gamma (IFN-gamma) PE-Cy®7 conjugate (Cat. no. A18713) and anti-mouse tumor necrosis factor alpha (TNF-a) Pacific Blue™ conjugate (Cat. no. RM90128). Samples were collected using the Attune® Acoustic Focusing Cytometer (blue/violet) with 488 nm excitation and 530/30 nm bandpass emission filter to detect FITC, and 640 nm longpass filter to detect PE-Cy®7 conjugate. 405 nm excitation and 450/40 nm bandpass emission filter were used to detect the Pacific Blue™ fluorescence. PANEL A: IFN-gamma and TNF-a co-staining of total mouse splenocytes that were left unstimulated (left panel) or stimulated (right panel) with PMA and ionomycin in the presence of brefeldin A. PANEL B: CD4 T cell expression of TNF-a (left panel) and IFN-gamma (right panel) after stimulation with the above stated conditions.
|Tested species reactivity||Mouse|
|Host / Isotype||Rat / IgG1, kappa|
|Immunogen||E.coli-expressed recombinant mouse IFN gamma|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin|
|Contains||0.09% 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.
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.