Serum starved HepG2 cells were incubated at 37°C for 30 minutes with vehicle control (0 μM) and different concentrations of (±)-naringenin (ab120958). Increased expression of p38 (phospho T180 + Y182) (

Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10μg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with μg /ml and ab8227 at 1/1000 dilution overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP (ab97040) at 1/10000 and visualised using ECL development solution.

All lanes : Anti-p38 (phospho T180 + Y182) antibody [M139] (ab45381) at 1/1000 dilution

Lane 1 : A431 cells serum starved overnight
Lane 2 : A431 cells treated with pervanadate (1 mM) for 30 minutes
Lane 3 : A431 cells treated with pervanadate (1 mM) for 30 minutes with No peptide
Lane 4 : A431 cells treated with pervanadate (1 mM) for 30 minutes with phospho-ERK1 (T202/Y204) peptide
Lane 5 : A431 cells treated with pervanadate (1 mM) for 30 minutes with phoshpo-p38 (T180/Y182) peptide

Lysates/proteins at 20 µg per lane.

Predicted band size: 42 kDa
Observed band size: 42 kDa

Immunofluorescence analysis of pervanadate treated mouse C2C12 cells, staining p38 (phospho T180 + Y182) with ab45381 at 1/250 dilution. Cy3-conjugated anti-mouse IgG was used as the secondary antibody.