A carrier-free 32P-labeled Edman reagent for the detection of proteins in the femtomole range.
A new route for synthesis of a 32P-labeled Edman reagent 2-(4-isothiocyanatophenoxy)-1,3,2-dioxaphosphinane 2-oxide (PEPITC) is reported. The carrier-free [32P]-PEPITC (specific radioactivity 3.3 X 10(17) Bq/mol) in theory allows the labeling and detection of a minimum amount of 0.1 femtomol of protein. The advantages of the new reagent, which has previously been used for protein sequencing, are demonstrated by gelelectrophoretical and immunological experiments. Its major role is seen in the quantitative monitoring of protein elution from preparative one- and two-dimensional gels. Proteins thus obtained are, after the usual desalting steps, ready for sequencing.
D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007202:Indicators and Reagents; D008832:Microchemistry; D009943:Organophosphorus Compounds; D010761:Phosphorus Radioisotopes; D011506:Proteins / Q000032:analysis*; D011865:Radioisotope Dilution Technique
C043517:2-(4-isothiocyanatophenoxy)-1,3,2-dioxaphosphinene 2-oxide; D007202:Indicators and Reagents; D009943:Organophosphorus Compounds; D010761:Phosphorus Radioisotopes; D011506:Proteins