CelLytic P

Figure 1. Compatibility with Gel Shift Assay.
Protein extracts were prepared with the CelLytic P reagent from spinach leaves. A double stranded ³²P labeled CREB oligonucleotide probe was incubated with 28 µg of the whole cell extract (lanes 2-5) or without whole cell extract (lane 1, free probe). Binding reactions with the extracts were performed in the absence [–] of competitor oligonucleotide (lanes 1-2) or in the presence of 100 or 500 fold excess of unlabeled CREB binding motif oligonucleotide (specific competitor [SP], lane 3 and lane 4, respectively) or in the presence of 100 fold excess of unlabeled oligonucleotide (non specific competitor, [NS] lane 5). Binding reactions were run on a non-denaturing 6% polyacrylamide gel, dried, and imaged on X-ray film. The arrows indicate the CREB-DNA complex and the free probe.