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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 & 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|
|Immunogen||Mouse TNF Alpha.|
|Storage buffer||PBS with 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.
This gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism, and coagulation. This cytokine has been implicated in a variety of diseases, including autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested the neuroprotective function of this cytokine.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
APC1 protein; Cachectin; macrophage-derived; member 2; monocyte-derived; TNF; TNF superfamily; TNF-a; TNF-alpha; TNFA; Tumor necrosis factor; tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor-alpha
DIF; Tnf; TNF-a; TNF-alpha; Tnfa; TNFalpha; Tnfsf1a; TNFSF2