Anti-AKINB1 | SNF1-related protein kinase regulatory subunit beta-1

Arabidopsis thaliana

Application example

25 µg of empty vector (1), pCDNA 3.1 – HA- tagged AKIN-beta1 (2), pCDNA 3.1 – HA- tagged AKIN-beta2 (3), 50 µg of total protein from Arabidopsis thaliana leaves had been homogenized into 50 mM NaPO4 pH 7.5, 20 mM KCl, 0.5 M sucrose, 0.2 mM PMSF, 10 mM DTT and protease inhibitor cocktail (Sigma, P9599). Extracts were separared on 10%NuPage Bis-Tris Novex SDS PAGE (Invitrogen) gels followed by a transfer for 60 min to PVDF membrane. Filters were blocked for 1h with 5% low-fat milk powder in PBS-T (0.1% TWEEN 20) and probed with anti-AKINß 1 antibodies (2 µg/ml for 60 min at RT) followed by Protein G-HRP (Bio-Rad; 1:3 000 for 60 min at RT) in PBS-T. Antibody incubations were followed by washings in PBS-T (3 x 5 min). All washing steps were performed at RT with agitation. Signal was detected with ECL (Millipore) using CCD camera. Exposure time was 10 seconds.

Courtesy Dr. David Stapleton, Melbourne, Australia

AKIN beta-1 (AKINB1) is a member of SnRK1/SNF1/AMPK family in plants. It is a regulatory subunit of the putative trimeric SNF1-related protein kinase (SnRK) comples which may play a role in a signal transduction cascade regulating gene expression and carbohydrate metabolism in higher plants.