DEGP12 | Putative protease Do-like 12 (mitochondrial)

40, 20 and 10 µg of recombinant Arabidopsis thaliana DEG12 were separated on 8 % SDS-PAGE  and blotted 1h to PVDF using semi-dry transfer. Blot was blocked with 5 % milk for 2h/RT  with agitation. Blot was incubated in the primary antibody at a dilution of 3.4 µg/ml in PBS-T 1h/RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 4 times for 10 min. in PBS-T at RT with agitation. Blot was incubated in the matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:50 000 in  for 1h/RT with agitation. The blot was washed as above and developed with chemiluminescent detection reagent, following manufacture's instructions.