Quantitative in situ gel electrophoretic assay for neomycin phosphotransferase activity in mammalian cell lysates

Nevis Fregien, Norman Davidson

Research output: Contribution to journalArticle

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Abstract

A method for the assay of neomycin phosphotransferase activity in eucaryotic cell lysates is described. Total cytoplasmic proteins are fractionated in nondenaturing polyacrylamide gels and then allowed to react in situ with [γ-32P]ATP and kanamycin. The reaction products are detected by blotting to phosphocellulose paper and autoradiography. The assay is linear with protein concentration and sensitive enough to detect expression in transient assays.

Original languageEnglish (US)
Pages (from-to)101-104
Number of pages4
JournalAnalytical Biochemistry
Volume148
Issue number1
DOIs
StatePublished - Jul 1985

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Keywords

  • gene transfer
  • neomycin phosphotransferase
  • nondenaturing gel electrophoresis

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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