IP - How much protein needed? - (May/28/2006 )

Hi

I want to do an IP.
How much total protein is normally used to do an IP ?
How many mg?
I guess it depends on the protein expression levels and antibody used.
Can you please give some examples of the amounts you work with?
Thanks

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just to give you an idea
Usually it's 100 to 500 µg. It depends on the protein, the cell and the antibody
Once I went up to 800 µg (focal adhesion kinase from old fibroblasts)

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Hi Missele,

Thanks for your eg.
Just few wuestions about i?
You are working with non-transfected cells, right?
Is the protein you IP highly or low expressed?

Thanks

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non transfected cells.
not really high expression, but not the lowest.

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