Journal
ANALYTICAL BIOCHEMISTRY
Volume 503, Issue -, Pages 56-57Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2016.03.017
Keywords
LacZ; B-Galactosidase (Bgal); beta-Galactosidase; Microplate reader
Funding
- Biotechnology and Biological Sciences Research Council (BBSRC) [BB/J00717X/1]
- U.K. Medical Research Council (MRC) project grants [MR/M017672/1]
- BBSRC [BB/J00717X/1] Funding Source: UKRI
- MRC [MR/M017672/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/J00717X/1] Funding Source: researchfish
- Medical Research Council [MR/M017672/1] Funding Source: researchfish
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Historically, the lacZ gene is one of the most universally used reporters of gene expression in molecular biology. Its activity can be quantified using an artificial substrate, o-nitrophenyl-ss-D-galactopyranoside (ONPG). However, the traditional method for measuring LacZ activity (first described by J. H. Miller in 1972) can be challenging for a large number of samples, is prone to variability, and involves hazardous compounds for lysis (e.g., chloroform, toluene). Here we describe a single-step assay using a 96-well microplate reader with a proven alternative cell permeabilization method. This modified protocol reduces handling time by 90%. (C) 2016 The Authors. Published by Elsevier Inc.
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