Puzzled__How to identify the interaction of two protein! - I had disclosed FGF inhibited the X protein.But I can not explain the (Jun/27/2007 )
Hi there!
I had disclosed FGFs inhibited the X protein.But I can not explain the phenomenon from the singnal pathway.My advisor asked me to make a paper for the subsequent experiment!I am very confused,I can not find a way out to the next.I do not konw hwo to identify the interaction of two protein!Where is the point!
Only a protocol or a advize can be very helpful!
Co-immunoprecipitation would be one way (see other posts in this section or Immunology section). Other ways involved cloning and expressing protein X as fusion proteins (GST fusions and using pulldowns or in yeast 3-hybrid system as a bait).
Best wishes,
Ceri
I had disclosed FGFs inhibited the X protein.But I can not explain the phenomenon from the singnal pathway.My advisor asked me to make a paper for the subsequent experiment!I am very confused,I can not find a way out to the next.I do not konw hwo to identify the interaction of two protein!Where is the point!
Only a protocol or a advize can be very helpful!
what means "FGF inhibited the x protein"? is FGF fibroblast growth factor? how should it inhibit X protein?
FGF play important role indirectly with X protein, Co-immunoprecipitation can not evalute the associations between the two! Does any other application more efficent?
do you know whether X protein would be modified (such as phosphorylation, methylation) upon -/+ FGF treatment? Could you find specific antibody for X protein (not pan antibody)?
I had disclosed FGFs inhibited the X protein.But I can not explain the phenomenon from the singnal pathway.My advisor asked me to make a paper for the subsequent experiment!I am very confused,I can not find a way out to the next.I do not konw hwo to identify the interaction of two protein!Where is the point!
Only a protocol or a advize can be very helpful!
What is with microscopical techniques (colocalization studies)? or don´t you know the interacting protein?
I don´t know, but maybe you could try a chromatographic method: where protein x can bind to FGF (on the column).
I have confirmed that FGF did not bind with the X protein,X protein have been konwn to me.But the mechanism of antagonize function from FGF to X protein was unclear.Specially in the singnal pathway.I want to know some method ,which can rapidly explan or find the signal between the two protein .
thanks