Hi there,
I am in a genetics lab and do not have much experience in biochemistry and please excuse my naive question.
I am planning to look at if my protein A is stabilized by another protein B in cell culture system. So I am going to do a pulse-chase experiment to compare the turn-over rate of protein A in the presence and absence of protein B. I will transfect cells with either A or A+B, radioactively label them, replace the radioactive media and them harvest the cell at different time points after media replacement. Cell lysates will be immunoprecipitated with anti-A antibody. Then the half-life of protein A will be monitored by SDS-PAGE. So my question is: if my hypothesis is true (A is stabilized when B is bound to it), shall I always see more A in the presence of B compared to A alone? Since B will be radioactively labeled and co-immunoprecipitated with anti-A antibody. Thank you very much and look forward to everyone's reply.
