I just ran an in vitro kinase assay that produced perplexing
results. I immunoprecipitated my kinase from monocyte-derived
macrophages, gave it enolase as a substrate and hot ATP-gamma, and then
ran the results (which should be mostly labeled enolase) on a minigel. I
ran the free nucleotide off of the end of gel and then made an autorad.
There were no distinct bands, but each lane showed an equal distribution
of label, ie., the entire lane was one big band. Anyone have any clue
what happened?
