Co-IP from tissues - Co-IP protocol for tissues (Jun/22/2007 )

I everybody!!!

I would like to do Co-IP from tumor sample. Does anybody of you had some experience and could provide the protocol.

thank you in advance,

Karmen

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depends on preparation, type of Ab´s , and precipitation method; first, you have to choose the appropriate Ab´s, and then look for protocols specific to these Ab and your lysate/homogenate

your question is too general to be properly answered

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Hi Bearer!!!

Thank you for your reply. I'll be using polyclonal antibodies and precipitating them with protein G sepharose beads.
Have you ever Co-Iped from tissues? If so, which lysis buffer did you use???

thank you in advance,
Karmen

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tissues or mostly homogeneized, not lysed (beside you work with homogenous in vitro tissues);

as you normally do not work with detergents in homogeneization, you may subfractionate your homogenate

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Bearer, thank you!!!

How do you subfractionate???
Somebody told me to use teflon balls for homogeneization. Have you ever tried???

Karmen

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I have heard of it: it should give a better grinding of cells but I never tried

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