Immunoprecipitation Question - Is it Possible to pull down a receptor by probing for its ligand (Jan/23/2007 )
basically I need to know if the following is feasible. I am working with a peptide that has an unidentified receptor; would it be possible to IP the receptor from a homogenate that very likely expresses it by incubating the homogenate with the peptide, followed by an antibody for the peptide, and ending with typical IP procedures? Is it likely or even possible that the ligand-receptor interaction would hold up through the IP procedure?
I've never tried anything like this and neither have any of my colleagues so please excuse if this is a ridiculous question.
Thanks in advance.
-Mondo977-
QUOTE (Mondo977 @ Jan 23 2007, 09:28 AM)
basically I need to know if the following is feasible. I am working with a peptide that has an unidentified receptor; would it be possible to IP the receptor from a homogenate that very likely expresses it by incubating the homogenate with the peptide, followed by an antibody for the peptide, and ending with typical IP procedures? Is it likely or even possible that the ligand-receptor interaction would hold up through the IP procedure?
I've never tried anything like this and neither have any of my colleagues so please excuse if this is a ridiculous question.
Thanks in advance.
I've never tried anything like this and neither have any of my colleagues so please excuse if this is a ridiculous question.
Thanks in advance.
the best thing of course is to try and see what happens; if your peptide has a good affinity to the receptor, it may remain bound if the anti-receptor Ab is not directed against the peptide binding site and you chhoese the right buffer;
maybe think of immobilizing the peptide (to beads, f.i.) and try to fish the receptor by binding to the peptide; as you like to use several peptides you will be faced with a lot unspecific binding effects; my proposal may diminish this unspecific background
-The Bearer-