Anti-CBP | CREB-binding protein homolog

application example


Total nuclear protein from 2x10^6 Drosophila melanogaster cultured cells was separated on 6% SDS-PAGE and blotted to PVDF using tank transfer. The blot was incubated in the primary antibody at a dilution of 1: 500 in 1% BSA, 0.05% Tween-20, 1x PBS for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice with 1x PBS. Blot was incubated in secondary antibody (anti-rabbit IgG AP-conjugated) diluted to 1:10 000 in 1% BSA, 0.05% Tween-20, 1x PBS for 30 min at RT with agitation. The blot was rinsed twice with 1x PBS and equilibrated in detection buffer (100mM NaCl; 10mM Tris-HCl pH9.5; 10mM MgCl2). The blot was incubated in substrate solution: 165ug/ml of BCIP and 330ug/ml of NBT in detection buffer until the signal reached the desired contrast and then rinsed with water and air dried.

Cortesy of Dr. Tatyana Kahn, Umeå University, Sweden