Question

Why is radioactive ATP bad for western blots from a kinase assay

Answer 1

Radioactive ATP in Western Blot for kinase assay is not a good experimental model because

All experiments with radioactive ATP should be done in a particular place for radioactive experiments. ATP storage should be in -20 degree centigrade and specific container for freezing and thawing is needed.

The gel with protein and buffer will be containing the radioisotope and needed to be discarded after use.

Even after the in vitro kinase assay, the experiment should be proved in vivo system.

Chances of multiple kinase-mediated pathways cannot be determined using autoradiography. Only site-specific phosphorylation can be done with radioactive ATP. Rather, the use of non-radioactive process may describe the site of phosphorylation.