Anti-Vimentin (phospho Ser39) Antibody (A90575)

Western blot analysis of extracts of NIH/3T3 cells, using Anti-Vimentin (phospho Ser39) Antibody (A90575) at 1:1,000 dilution. NIH/3T3 cells were treated by Paclitaxel (100 nM/ml) at 37°C for 20 hours. NIH/3T3 cells were treated by Hydroxyurea (4 mM) at 37°C for 20 hours. 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 Enhanced Kit (RM00021). Exposure time: 1s.

Western blot analysis of extracts of HeLa cells, using Anti-Vimentin (phospho Ser39) Antibody (A90575) at 1:1,000 dilution. HeLa cells were treated by Paclitaxel (100 nM/ml) at 37°C for 20 hours. HeLa cells were treated by Hydroxyurea (4 mM) at 37°C for 20 hours. 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: 180s.

Immunohistochemistry analysis of paraffin-embedded human appendix tissue using Anti-Vimentin (phospho Ser39) Antibody (A90575) at a dilution of 1:100 (40x lens). Perform microwave antigen retrieval with 10 mM Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.