Anti-BAK1 | Brassinosteroid insensitive 1-associated receptor kinase 1 (monocots)

µg of total protein from    extracted with write exact buffer components ..... and denatured with....at C or 70C for 2-5 min.... were separated on  % SDS-PAGE  and blotted 1h to PVDF using semi-dry or tank transfer. Blots were blocked with   for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT 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 TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from ) diluted to 1:0 000 in  for 1h at RT with agitation. The blot was washed as above and developed for   min with   EXACT NAME OF REAGENT AND SUPPLIER. Exposure time was  seconds.

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