ACC synthase 8 | 1-aminocyclopropane-1-carboxylate synthase 8

80 and 40 µg of recombinant ACS8 protein were separated on  8 % SDS-PAGE  and blotted 1h to PVDF. Blot was blocked with 5 % milk for 2h/RT in PBS-T with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h/RT with agitation in PBS-T or ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 4x in PBS-T at RT with agitation. Blot was incubated in 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 according to manufacture's instructions.