Enzyme activity dot blots: a rapid and convenient assay for acetyltransferase or protein kinase activity immobilized on nitrocellulose

Anal Biochem. 1986 Nov 15;159(1):58-66. doi: 10.1016/0003-2697(86)90307-6.

Abstract

Methods are described for assaying (Tetrahymena) histone acetyltransferase activity and (Drosophila) casein kinase II activity by spotting extracts on nitrocellulose filters. The methods are quantitative over a wide range of enzyme concentrations and are almost as sensitive as liquid assays. Examples are presented for illustrating the use of these methods for enzyme purification, concentration, and desalting, as well as for electrophoretic blotting from agarose gels. A simple method for autoradiographic enhancement of nitrocellulose filters is also described.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acetyltransferases / analysis*
  • Animals
  • Casein Kinases
  • Collodion
  • Drosophila / enzymology
  • Electrophoresis
  • Enzymes, Immobilized*
  • Histone Acetyltransferases
  • Protein Kinases / analysis*
  • Saccharomyces cerevisiae Proteins*
  • Tetrahymena / enzymology

Substances

  • Enzymes, Immobilized
  • Saccharomyces cerevisiae Proteins
  • Collodion
  • Acetyltransferases
  • Histone Acetyltransferases
  • Protein Kinases
  • Casein Kinases