Application
Western blot
We performed an IP against Bim from HCT116 cell lysate. As the protein size of Bim is around 25 kDa, we first tested the VeriBlot secondary antibody (ab131366) to analyse its binding (lane 1). No signal from the IgG heavy and light chain, even after long exposure times, was observed when using ab131366. Next we developed the same membrane with a standard secondary antibody (ab97051) as a comparison (lane 2). The signal was much stronger with the standard secondary antibody but the strong signals of heavy and light chain of IgG overlayed the signal of Bim. To sum up, the VeriBlot secondary antibody is a nice tool to analyse proteins at around 25 or 55 kDa in IPs.
Lane 1 shows Bim long and short forms, after a exposure time of 5 min with ECL+. The secondary antibody (ab131366) was diluted 1:2000. Lane 2 shows Bim overlayed by the light chain of IgG after a exposure time of 2 min with ECL. The secondary antibody (ab97051) was diluted 1:10000.
Lane 1 shows Bim long and short forms, after a exposure time of 5 min with ECL+. The secondary antibody (ab131366) was diluted 1:2000. Lane 2 shows Bim overlayed by the light chain of IgG after a exposure time of 2 min with ECL. The secondary antibody (ab97051) was diluted 1:10000.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
MR. Christian Marx
Verified customer
投稿 Aug 27 2013