Hello
I am planning to do a quality check of my antibodies to different GABAA receptor via immunocytochemistry? I have a1, b2 and gamma2s subunits. If I want to verify that the b2 antibody binds to b2 and not other subunits, how do I do it? Do I need to transfect a different combination like a6 b2 delta subunit (for example)?
It is grateful for me to hear other people's ideas and experiences.
Hoping for your insightful answers.
The easiest way, as long as your antibody is good for both IHC and western blot is to perform western blot analysis.
I'd overexpress the proteins in question and look for differences in expression levels by IHC and WB. Alternatively, try immunoprecipitation and analyze what has been captured. This gives you the option to positively identify the proteins by MS.
The cleanest way is to use CRISPR to remove the subunit of interest. An IHC on WT and KO will show if your Ab binds to other subunits or not. A control Ab that binds to other subunits will be a good addition to the experiment.
- Badges
- Science topic
- Similar topics
- Mathematics