GH | Gamma-glutamyl hydrolase

Various amounts of Glycine max recombinant GH were loaded/well and separated on 8 % SDS-PAGE  and blotted 1h to PVDF. Blot was blocked with 5 % milk for 2h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 2.7 µg/ml for 1h/RT with agitation in PBS-T 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 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, following manufacture's recommendation.