Western blotting…*sigh*…I have been performing Western blots on my peptides which are labelled with both a GST tag and a HIS tag on their N- and C-terminal ends respectively. Whenever I perform the blot with an anti-GST antibody, it works perfectly, yet whenever I perform the blot with the anti-HIS antibody, it never works properly. My samples would never appear, and my controls would be faint. I cleaved the GST tag from my peptides (with Thrombin, stopping the reaction with PMSF) and then performed a blot with the anti-HIS antibody again, and now nothing shows up, only non-specific blotches all over the exposed film!
If anyone has any suggestions as to what might be interfering with it, I’d greatly appreciate it. Thanks Image may be NSFW.
Clik here to view.
FYI, I have also posted this discussion on Nature Network.
http://network.nature.com/groups/labvamp/forum/topics
If anyone has any suggestions as to what might be interfering with it, I’d greatly appreciate it. Thanks Image may be NSFW.
Clik here to view.
FYI, I have also posted this discussion on Nature Network.
http://network.nature.com/groups/labvamp/forum/topics
