Rabbit anti p38 mapk antibody

Proteins were measured as previously described [4 (link), 6 (link), 9 (link)] using mouse anti-CBS (A-2) antibody, a mouse anti-CSE (30.7) antibody, mouse anti-MPST (H-11), rabbit anti-p38 MAPK antibody and rabbit anti–phospho–p38 MAPK (Thr180/Tyr182) antibody (all from Santa Cruz Biotechnology, Middlesex, UK) and, rabbit anti–extracellular signal–regulated kinase (ERK)–1/2 (137F5) and rabbit anti–phospho–ERK-1/2 (Thr202/Tyr204; purchased from Cell Signalling Technology, Ely, Cambridgeshire, UK).

For whole-cell extract preparation, nucleated red blood cell-enriched mononuclear cells were lysed in radioimmunoprecipitation assay (RIPA) buffer containing a complete protease inhibitor cocktail (Roche, Penzberg, Upper Bavaria, Germany) for 10 min at 4°C and the extract (50 μg) was separated using SDS-PAGE and transferred onto polyvinyl difluoride (PVDF) membranes. Membranes were incubated with rabbit anti-p38 MAPK antibody (1 : 500), goat anti-mouse β-actin antibody (Santa Cruz Biotechnology, Santa Cruz, CA), or rabbit anti-phospho-p38 MAPK antibody (1 : 500, Bioworld Technology, St. Louis Park, MN). HRP-conjugated anti-goat or anti-rabbit secondary antibodies (both 1 : 5000) were purchased from Sigma Aldrich (St. Louis, MO) and Santa Cruz Biotechnology, respectively. The sample sizes pertaining to the Western blot analyses for the three groups were similar to those described for previous experiments; that is, 13 patients treated with R. Astragali, 11 patients treated with the compounded formulation, and 11 patients treated with the placebo drug were analyzed during the Western blot analyses.