COP1 | constitutive photomorphogenesis protein 1

3.5 µg of total protein from 4-day old Vicia faba roots, was separated on 12 % SDS-PAGE and blotted 50 min to PVDF membrane. Blots were blocked immediately following transfer in MTBS-T (5 % milk) for 30 min. at RT with agitation. Blots were incubated in the primary antibody at a dilution of 1: 2000 for 1 h ar RT with agitation. The antibody solution ws decanted and the blot was rinsed briefly twice, following by a 3 times was for 3 min. in TBS-T at RT with agitation. Blots were incubated with secondary antibody (anti-rabbit IgG, HRP conjugated from Agrisera, AS09 602) in 1: 20 000 dilution for 30 min. at RT with agitation. The blots were washed as above and developed for 5 min. with ECL Advance detection reagent (GE Healthcare) according to the manufactures instructions. Exposure time was 60 seconds.

Courtesy of Dr. Dorota Rybaczek

Arabidopsis thaliana plants have been cultured for 5 days in darkness. Immunolocalization was done according to a standard protocol. Primary antibody anti-COP1 was applied in 1:400 dilution, (shown in green) anti-PIN1 mous monoclonal antibody (shown in red) and a secondary antibody anti-mouse 555 and anti-rabbit 488 (Alexa), diluted 1: 500. Scale bar- 20 µm.

Courtesy of Dr. Taras Pasternak, Freiburg University, Germany.