Quantification of subnanomolar amounts of phosphate bound to seryl and threonyl residues in phosphoproteins using alkaline hydrolysis and malachite green.
An assay for protein-bound phosphate with a capacity to determine 100 pmol of phosphate is described. It is based on the combination of two well known methods: the alkaline hydrolysis of phosphate from seryl and threonyl residues in phosphoproteins and the quantification of the released phosphate by the use of malachite green and phosphomolybdate.
Ekman
P
P
Department of Medical and Physiological Chemistry, Biomedical Center, Uppsala, Sweden.
Jäger
O
O
D000255:Adenosine Triphosphate / Q000378:metabolism; D002364:Caseins / Q000737:chemistry; D004396:Coloring Agents; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D005340:Fibrinogen / Q000737:chemistry; D006657:Histones / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008832:Microchemistry / Q000379:methods; D010047:Ovalbumin / Q000737:chemistry; D010710:Phosphates / Q000032:analysis*; D010750:Phosphoproteins / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D010768:Phosphoserine / Q000032:analysis*; D010769:Phosphothreonine / Q000032:analysis*; D011479:Protamines / Q000737:chemistry; D012394:Rosaniline Dyes; D013053:Spectrophotometry / Q000379:methods
D002364:Caseins; D004396:Coloring Agents; D006657:Histones; D010710:Phosphates; D010750:Phosphoproteins; D011479:Protamines; D012394:Rosaniline Dyes; D010769:Phosphothreonine; C005095:malachite green; D010768:Phosphoserine; D000255:Adenosine Triphosphate; D005340:Fibrinogen; D010047:Ovalbumin; D017868:Cyclic AMP-Dependent Protein Kinases