Phosphatase activity after IP - (Mar/27/2007 )
Hi all,
i have some metabolic evidences that my protein of interest (a cytosolic lipid transporter) interacts with a protein phosphatase.
My aim is to identify the protein phosphatase, using a co-IP approach. In order to restrict the number of potential protein phosphatases involved (PP1, PP2, etc), i would assay the phosphatase activity (if any) of the immunoprecipitate in the presence of various phosphatase inhibitor.
In my lab there is a working protocol for immunoprecipitate the lipid transporter and also a sensible, fluorogenic substrate for testing protein phosphatase activity.
My question is: is it possible to co-IP the potential protein phosphatase without disrupting its enzimatic activity?
Thanks in advance.
Thomas
i have some metabolic evidences that my protein of interest (a cytosolic lipid transporter) interacts with a protein phosphatase.
My aim is to identify the protein phosphatase, using a co-IP approach. In order to restrict the number of potential protein phosphatases involved (PP1, PP2, etc), i would assay the phosphatase activity (if any) of the immunoprecipitate in the presence of various phosphatase inhibitor.
In my lab there is a working protocol for immunoprecipitate the lipid transporter and also a sensible, fluorogenic substrate for testing protein phosphatase activity.
My question is: is it possible to co-IP the potential protein phosphatase without disrupting its enzimatic activity?
Thanks in advance.
Thomas
of course, if your Ab does not target and therefore blocks the catalytic domain of your phosphatase(s); do you use anti-PPP Ab´s?
I am interested in good working anti-PPP Ab´s (PP1, PP2A, PP2B, PTP1B) and need some recommendations
I'm going to co-IP protein phosphatase using an antibody against my protein of interest. So i can't help you, i'm sorry ;-)