AtpF | CF0I subunit of ATP synthase
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20 µg of chloroplast fraction from Arabidopsis thaliana (1) and Spinacia oleracea (2) were separated on 12 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with 2 % non-fat milk powder in 1xTBS-T for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions.
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