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Anti-IL27 Antibody 3H12.F10
Also for IL27 (NM_145636)
|This Protein A purified monoclonal antibody was produced in rats by repeated immunizations with mature length recombinant mouse p28 protein (produced in E.coli) followed by hybridoma development.|
||ELISA: 1:10,000, WB: 1:1000
|0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2|
|Mouse IL-27/p28 Subunit, also known as Interleukin-30, is a member of the IL-12 family of cytokines. When combined with EBI3 (Epstein-Barr virus induced gene 3), the heterodimer formed is IL-27. Mouse p28 is a proinflammatory cytokine inducing immunomodulatory effects. Current research is underway to delineate specific biological functions.
|Mus musculus interleukin 27 (Il27)|
|IL-27; IL-27p28; Il30; p28|
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Anti-IL-27/p28 antibody in western blot shows detection of recombinant mouse IL-27/p28. Recombinant protein (0.1 µg) was loaded on to an SDS-PAGE gel, and after separation, transferred to nitrocellulose. The expected band is approximately 26 kDa in size. The membrane was blocked with 1% BSA in TBST for 30 min at RT, followed by incubation with Anti-IL-27/p28 antibody diluted 1:1,000 in 1% BSA in TBST overnight at 4°C. After washes, the blot was reacted with secondary antibody HRP Goat anti-Rabbit IgG antibody diluted 1:40,000 in blocking buffer (p/n MB-070) for 30 min at RT. Data was collected using Bio-Rad VersaDoc® 4000 MP imaging system.
Mouse peritoneal macrophages were grown in culture for 24 hours, stimulated with 10ng/mL IFN? and 1ug/mL LPS for 14 hours and incubated for 4 hours with Bredfeldin A. Cells were harvested, washed, aliquoted 1x106 cells per sample, and fixed and permeabilized according to a standard protocol. Samples were stained with biotinylated primary anti-mouse p28 antibody at (0.1 - 10ug/mL primary antibody alongside negative controls of unstimulated cells and isotype controls. Cells were stained with 0.25ug/mL rat anti-mouse CD107b conjugated Alexa Fluor 647 and PHYCOERYTHRIN Conjugated secondary at 1:100 and analyzed by flow cytometry. Stimulated cells showed increase PE staining (horizontal axis) when compared with unstimulated cells.