1 reply to this topic

[gaiteruprotocol]

Hi everybody. I´m planning on starting to study in vivo phosphorylation of my protein. As a first tentative, I thought on transiently expressing constructs of my protein, immunoprecipitating them from lysates, and trying to assess any phosphorylation using ProQ Diamond (Molecular Probes) as we don't have any phosphospecific antobody and we don't know which aminoacid might be the target of the modification. Has anybody had experience with this reagent? Is it sensitive enough for using it in this approach?

Thanks a lot in advance and a good start of the week.

[linyjing]

gaiteruprotocol, on May 4 2009, 08:31 AM, said:

Hi everybody. I´m planning on starting to study in vivo phosphorylation of my protein. As a first tentative, I thought on transiently expressing constructs of my protein, immunoprecipitating them from lysates, and trying to assess any phosphorylation using ProQ Diamond (Molecular Probes) as we don't have any phosphospecific antobody and we don't know which aminoacid might be the target of the modification. Has anybody had experience with this reagent? Is it sensitive enough for using it in this approach?

Thanks a lot in advance and a good start of the week.

hi, I used ProQ to stain my protein after I did in vitro kinase assay. I used 1 ug protein as substrate, and its phosphorylated form was stained. not sure whether it is sensitive enough for your experiment. have you tried serine/threonine/tyrosine phoshorylation ab?