Anti-PBCP | Photosystem II core phosphatase

Protein extracts were denatured with 5X SDS buffer diluted to 1X (iNtRON, biotechnology), the exact buffer components:Tris-HCl pH 6.8 250 mM, SDS 10%, Glycerol 50%, DTT 0.5M, bromophenol blue 0.5% incubated at room temperature (RT) for 30 mins. Samples were separated in the cold on 10 % SDS-PAGE and blotted for1 h to PVDF memrane (pore size of 0.45 um), using: wet transfer in the cold. Blot was blocked with 5 % skim milk for 1h/RT with agitation. Blot was incubated in the primary antibody (AS23-4991) at a dilution of 1: 2000 1hr/RT in TBST with agitation. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed 4 time for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 25 000 in for 2h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent.

Courtesy of Dr. Khan Naveed, Pusan National University, Korea

Alternative name: AtPP2C26