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

Nevis Fregien; Norman Davidson

doi:10.1016/0003-2697(85)90633-5

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

Nevis Fregien, Norman Davidson

Research output: Contribution to journal › Article › peer-review

9 Scopus citations

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 language	English (US)
Pages (from-to)	101-104
Number of pages	4
Journal	Analytical Biochemistry
Volume	148
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(85)90633-5
State	Published - Jul 1985
Externally published	Yes

Keywords

gene transfer
neomycin phosphotransferase
nondenaturing gel electrophoresis

ASJC Scopus subject areas

Biochemistry
Biophysics
Molecular Biology

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10.1016/0003-2697(85)90633-5

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title = "Quantitative in situ gel electrophoretic assay for neomycin phosphotransferase activity in mammalian cell lysates",

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.",

keywords = "gene transfer, neomycin phosphotransferase, nondenaturing gel electrophoresis",

author = "Nevis Fregien and Norman Davidson",

note = "Funding Information: This work was supported by a grant from Amgen Corporation.",

year = "1985",

month = jul,

doi = "10.1016/0003-2697(85)90633-5",

language = "English (US)",

volume = "148",

pages = "101--104",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Academic Press Inc.",

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TY - JOUR

T1 - Quantitative in situ gel electrophoretic assay for neomycin phosphotransferase activity in mammalian cell lysates

AU - Fregien, Nevis

AU - Davidson, Norman

N1 - Funding Information: This work was supported by a grant from Amgen Corporation.

PY - 1985/7

Y1 - 1985/7

N2 - 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.

AB - 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.

KW - gene transfer

KW - neomycin phosphotransferase

KW - nondenaturing gel electrophoresis

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U2 - 10.1016/0003-2697(85)90633-5

DO - 10.1016/0003-2697(85)90633-5

M3 - Article

C2 - 2994516

AN - SCOPUS:0021869143

VL - 148

SP - 101

EP - 104

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 1

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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