Anti-RPS12 | Ribosomal protein S12 (chloroplastic)

10 µg of total protein from Chlamydomonas reinhardtii extracted with standard lysis buffer (Tris-HCl pH 6.8 50mM, 10mM EDTA, 2% SDS, Sigma protease inhibitors)  were separated on 15 % SDS-PAGE and blotted to nitrocellulose membrane using a wet transfer cell. Blot was blocked in TBS-T containing 5% non-fat dry milk  for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 in TBS-T containing 5% non-fat dry milk  for 1h at RT with agitation. The antibody solution was decanted and the blot was washed  3 times for 15 min  in TBS-T containing 1% non-fat dry milk with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:0 000 in TBS-T containing 1% non-fat dry milk  for 1h at RT with agitation. The blot was washed as above  and developed for 1 min with ECL according to the manufacturers instructions. Exposure time was 30 seconds.