AS15 2859 Anti-AKINB3 | SNF1-related protein kinase regulatory subunit beta-3

Recombinant AKIN beta 3 were separated by 10% SDS-PAGE and transferred to a PVDF membrane using semi-dry blotting (for 1h). Blots were blocked with 5% low-fat milk powder in TBS-T (0.01% Tween 20) for 1h at room temperature (RT) with agitation. The blot was then incubated overnight at 4 °C in the primary antibody at a dilution of 1: 1 000 in 1% low-fat milk powder in TBS-T with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed 3 times for 5 min in TBS-T at RT with agitation. The blot was then incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera) diluted to 1:10 000 in 1% low-fat milk powder in TBS-T for 1h at RT with agitation. The blot was washed 5 times for 5 min and developed for 2 min with chemiluminescence detection reagent in low picogram range, according to the manufacturer's instructions. Exposure time to film was 10 min.

Courtesy of PhD student Sander Hulsmans, KU Leuven, Belgium

AKIN ß3 (SNF1-related protein kinase regulatory subunit beta-3) is a regulatory subunit of the probable trimeric SNF1-related protein kinase (SnRK) complex, which may play a role in a signal transduction cascade regulating gene expression and carbohydrate metabolism in higher plants.