I would like to learn my students how to perform a co-immunoprecipitation experiment, coupled to western blot of primary antigen and the co-precipitant.
Does anyone have a good suggestion for this? Failsave and not to expensive?
It's possible to do transfection for this purpose and precipitate using a tag antibody.
I may suggest to pull a down a cell membrane receptor that has different subunit such as integrin, I also recommend use of fast growing cells such as 293. For example, anti-integrin alpha V that has Mw >135 kDa, after Co-IP you may stain with anti-integrin beta 1 that has around 130 kDa.
But also if a transfection can be done, maybe myc-flag may fulfill this role, therefore a anti-myc and anti-flag can be used to analyze.
Best
I've done this as a discussion activity just using colored markers & paper.
You can consider the setup from this paper by my friend:
doi: 10.18632/oncotarget.18899
She performed mutant p53 - Hsp70 co-IP; this interaction is very strong. DO1 antibody for p53 IP is great. HSP70 is also easy to detect. You may overexpress proteins or not - depends on which cell line you work.
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