kinase assay - ATP (Feb/21/2008 )

Hi I did a kinase assay and it didn't work

I wondering if the ATP solution was not good since the ph was pretty acid (4.5)
even when I prepare in PBS.

how can i bring it back to 7, Naoh?



does it matter for kinase assay?

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pH of the assay buffer is important. Try to have something where the kinase is happy. And make up tris with pH 7.4 or 8.0 and try it out. The actual amount of ATP one might add is probably very small to alter the pH drastically. Thats why you need to have a good buffer for the assay.

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ok so my kinase buffer was ph 4.5.
is NaOH ok to bring it at ph7?



does it matter for kinase assay?[/quote]

pH of the assay buffer is important. Try to have something where the kinase is happy. And make up tris with pH 7.4 or 8.0 and try it out. The actual amount of ATP one might add is probably very small to alter the pH drastically. Thats why you need to have a good buffer for the assay.
[/quote]

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yes, use naoh to adjust the buffer.

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