Disadvantages of immunochemistry. - (Oct/26/2005 )

Question from my humble ignorance.

I want to determinate the quantity of a subunit which assembles to other three subunits. If I use immunochemistry, can I do it? Will I get interference because the link between the different subunits? How reliable is this method? Could you tell me the main disadvantages and probles of this technique in order to get good results (for ex. co-immunoprecipitation)?

Thanks you all in advance.

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I don't have time at the moment to go into exact details, however it's my opinion that immunocytochemistry is at best semiquantitaive. You would need to have controls w/known protein amounts, you would need to know the quantum output of the fluophore you use among other things.

For coIPs it is also at best semiquantitative, however I believe that Westerns are far easier to quantifiy than signal from cells/tissue. A simple standard curve repeated three times would be helpful. In any event you should run your samples on the same gel as your standard curve if you do any sort of scanning densitometry.

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