Anti-ERK1 (phospho Thr202) + ERK2 (phospho Thr185) Antibody (A16535)

Western blot analysis of extracts of various cell lines, using Anti-ERK1 (phospho Thr202) + ERK2 (phospho Thr185) Antibody (A16535) at 1:1,000 dilution. Jurkat and NIH/3T3 cells were treated by PMA/TPA (200 nM) at 37°C for 30 minutes after serum-starvation overnight. Jurkat cells were treated by ATP(5 mM) at 30°C for 1 hour. The secondary antibody was Goat Anti-Rabbit IgG H&L Antibody (HRP) at 1:10,000 dilution. Lysates/proteins were present at 25µg per lane. The blocking buffer used was 3% non-fat dry milk in TBST. Detection was with a ECL Basic Kit. Exposure time: 1s.

Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue using Anti-ERK1 (phospho Thr202) + ERK2 (phospho Thr185) Antibody (A16535). Perform microwave antigen retrieval with 10 mM Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Immunofluorescence analysis of methanol-fixed HeLa cells showing cytoplasmic, nuclear staining using Anti-ERK1 (phospho Thr202) + ERK2 (phospho Thr185) Antibody (A16535).