Immunoprecipitations were performed and samples were incubated at 95°C for 5 minutes in sample buffer containing SDS and beta-mercaptoethanol to fully reduce proteins. Samples were run on an SDS-PAGE gel and Western blots were performed, using Veriblot as the secondary antibody.
There were two IP samples.
Lane 1 - protein target MW: 75 kDa
Lane 2 - protein target MW: 22 kDa.
For sample 2, which IP'ed very well, the heavy chain signal was very low. For sample 1, however, which did not IP well, the heavy chain signal was very high. The asterisks mark the IP'ed protein. In both cases the light chain signal was very low or non-existent.
For proteins that do not IP well, are low abundance, or have poor primary antibodies, expect to see the heavy chain band. In those cases, as long as your protein of interest is not at ~50 kDa this product will work. If your protein is ~50 kDa, I would recommend using a secondary antibody that only recognizes the light chain (~25 kDa) instead.
There were two IP samples.
Lane 1 - protein target MW: 75 kDa
Lane 2 - protein target MW: 22 kDa.
For sample 2, which IP'ed very well, the heavy chain signal was very low. For sample 1, however, which did not IP well, the heavy chain signal was very high. The asterisks mark the IP'ed protein. In both cases the light chain signal was very low or non-existent.
For proteins that do not IP well, are low abundance, or have poor primary antibodies, expect to see the heavy chain band. In those cases, as long as your protein of interest is not at ~50 kDa this product will work. If your protein is ~50 kDa, I would recommend using a secondary antibody that only recognizes the light chain (~25 kDa) instead.
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.
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投稿 Feb 12 2021