soymilk14

i see. thank you. i have another last question sorry... if i have two cell lysates: no treatment and treatment groups. Can i use IP to distingusih if my target protein can bind to my IP antibody in no treatment group and can not bind in the treatment group? is that the purpose of using non denaturing buffers for IP? sorry about this, i hope you can clarify for me. thank you

oh i see, thank you Curtis. yes, the IP antibody i used is from rabbit and the western primary antibody i used is raised in mouse. haha thank you.  does that mean that my  western signal of my target protein is not true IP? does that mean that i got false positive, that my target protein really does not bind (directly or indirectly) to my IP protein? sorry for this, my IP concept is not strong. i hope you can help me clarify this or suggest a good reading material for IP basic concept.

another thing is my third question:

3. if  my target protein binds to protein A in the presence of a ligand (group A) , but does not bind in the absence of a ligand (group , if i do immunoprecipitation, can i still see this band in the group B? sorry i am confused on the concept, i might be misleading myself. i hope you can help me clarify this...


thank you so much