Phosphorylation specific p38 mapk antibody

The MAPK assays were described previously with some modifications (16 (link)). Briefly, rice suspension cells were subcultured in fresh medium for 3 days. Aliquots of 500 mg of cells were suspended in 5 mL preincubation medium (3% sucrose and 10 mM MES [morpholineethanesulfonic acid] in 5% culture medium [pH 5.8]) and incubated under culture conditions for 4 h. Total proteins were extracted with plant radioimmunoprecipitation assay (RIPA) lysis buffer (P0045; Beyotime Biotechnology, Shanghai, China) and phosphatase inhibitor cocktail (B15001; Bimake, Shanghai, China) from rice suspension cells subjected to treatment with ShAM1 or ShAM1-digested cell wall extract. Analysis was carried out by SDS–PAGE and Western blotting using a phosphorylation-specific p38 MAPK antibody (1:5,000 dilution) (Cell Signaling).

The MAPK assays were described previously with some modifications (16 (link)). Briefly, rice suspension cells were subcultured in fresh medium for 3 days. Aliquots of 500 mg of cells were suspended in 5 mL preincubation medium (3% sucrose and 10 mM MES [morpholineethanesulfonic acid] in 5% culture medium [pH 5.8]) and incubated under culture conditions for 4 h. Total proteins were extracted with plant radioimmunoprecipitation assay (RIPA) lysis buffer (P0045; Beyotime Biotechnology, Shanghai, China) and phosphatase inhibitor cocktail (B15001; Bimake, Shanghai, China) from rice suspension cells subjected to treatment with ShAM1 or ShAM1-digested cell wall extract. Analysis was carried out by SDS–PAGE and Western blotting using a phosphorylation-specific p38 MAPK antibody (1:5,000 dilution) (Cell Signaling).