Determination and occurrence of tyrosine O-sulfate in proteins

摘要: Publisher Summary This chapter focuses on the detection of tyrosine-sulfated proteins and determination tyrosine sulfate by methods that are based labeling with 35 SO 4 . approach has several advantages over chemical in unlabeled proteins. It can be much more easily used to search for is considerably sensitive, therefore requires less protein material analysis. The discusses an immunological detect In this approach, antisera were raised against a synthetic antigen containing large number residues antibodies purified from affinity chromatography. These appeared recognize sulfate-containing “Western” blots solid-phase radioimmunoassay. general usefulness these screen for, immunoprecipitate, or purify currently being tested. standard method involves four main steps: (1) vivo tissues situ tissue explants, slices, cells culture inorganic [ S]sulfate; (2) separation polyacrylamide gel electrophoresis (PAGE); (3) elution gels hydrolysis individual 4- 1abeled proteins; (4) S]sulfate thin-layer electrophoresis.