I would like to detect pyrophosphorylation in protein. Can you please tell me how I can detect pyrophosphorylation in protein? Is there any antibody available against the pyrophosphate?
many things can be done but roughly to begin with:
1. there are phospho antibodies, which can detect.
2. various mass spec methods can be used to detect phosphorylation.
3. if you have antibody against your protein, feed cells with radioactive phosphate and IP with antibody and autoradiogram.
I don't know exactly what you have that needs to be detected.
Rest is the paper above and many more papers available.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2000531/...... i imagine this would be helpful
many things can be done but roughly to begin with:
1. there are phospho antibodies, which can detect.
2. various mass spec methods can be used to detect phosphorylation.
3. if you have antibody against your protein, feed cells with radioactive phosphate and IP with antibody and autoradiogram.
I don't know exactly what you have that needs to be detected.
Rest is the paper above and many more papers available.
I am working as an undergraduate project student in the lab of Rashna Bhandari (whose paper Akhilesh has linked here).
Dorothea Fiedler (Princeton Uni) has been working on characterization and detection of artificially pyrophosphorylated polypeptides for a while and has recently come up with a reagent to specifically capture pyrophosphate group and has successfully demonstrated the capture of several arbitrarily designed pyrophosphorylated polypeptides.
Here is an early view of that paper:
http://onlinelibrary.wiley.com/doi/10.1002/anie.201411232/pdf
I don't think it will be commercially available very soon but the paper is indeed an interesting read and promising except the elimination of one or two possible non specific binders. Also look at the previous papers where they have given a detailed account of chemical nature of a pyrophosphopeptide.
Good luck understanding the most specific PTM ever !
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