Anti-PsbO | Photosystem II manganese-stabilizing polypeptide (algal)

Courtesy of Dr. David Vinyard, Louisiana State University, USA



10 µg/well of total protein extracted from Chlamydomonas reinhardtii, total cell extract were denatured with Invitrogen LDS sample buffer (4X) at 70°C/5 min. and separated on Invitrogen NuPage Bis-Tris 4-12% SDS-PAGE 45 min and blotted for 1 h to Invitrogen PVDF (pore size of 0.45 µm), using: wet transfer. Blot was blocked with 5% milk in TBS-T for: 1h/RT with agitation, and incubated in the primary antibody at a dilution of 1: 1000 for 1h/RT with TBS-T Blocking. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed once for 10 min and 2 times for 5 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG ALP conjugated, AS09 607 lot 2105 ) diluted to 1: 5 000 in TBS-T Blocking for 0,5h/RT with agitation. The blot was washed as above and developed with Agrisera AS19 BCIP-NBT reagent lot 03068231 for 3 minutes.