ArticleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society2016
Comments on Methods to Suppress Endogenous β-Galactosidase Activity in Mouse Tissues Expressing the LacZ Reporter Gene.
Article in The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 15 citations in OpenAlex.
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- Generation and characterization of a mouse model of conditional Chd4 knockout in the endometrial epithelium.PloS one · 2025Article
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- Reduced urine volume and changed renal sphingolipid metabolism in P2ry14-deficient mice.Frontiers in cell and developmental biology · 2023Article
- MCC is a centrosomal protein that relocalizes to non-centrosomal apical sites during intestinal cell differentiation.Journal of cell science · 2022Article
- Expression Pattern of theThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2021Article
- Article
- Tracing Gene Expression Through Detection of β-galactosidase Activity in Whole Mouse Embryos.Journal of visualized experiments : JoVE · 2018Article
- Absence of CFAP69 Causes Male Infertility due to Multiple Morphological Abnormalities of the Flagella in Human and Mouse.American journal of human genetics · 2018Article
- Advantages and Limitations of Salmon-Gal/Tetrazolium Salt Histochemistry for the Detection of LacZ Reporter Gene Activity in Murine Epithelial Tissue.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2017Article
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The Escherichia coli LacZ gene (encoding β-galactosidase) is a widely used reporter for gene regulation analysis in transgenic mice. Determination of β-galactosidase activity is classically performed using 5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside/ferri-/ferrocyanide (X-Gal/FeCN) histochemistry. Uncertainty about the origin of the β-galactosidase signal is encountered in tissues containing high levels of endogenous β-galactosidase. Here, we show that reliable results can nevertheless be obtained in these tissues by performing the histochemical reaction under slightly basic pH conditions (pH 8-9). We further demonstrate that in this context, analysis of tissue sections may be advantageous over that of conventional whole-mount tissues because poor dye penetration and remaining tissue acidity are avoided in tissue sections. We also recommend that bacterial debris should always be carefully removed from the luminal surface of gastrointestinal tract specimens unless staining of resident microflora is deliberately used as an internal positive control in the assay. Finally, we show that 6-chloro-3-indolyl-β-d-galactopyranoside with nitrotetrazolium blue chloride works well as an alternative chromogenic substrate for visualizing LacZ reporter gene expression in cryostat sections. Its use in high endogenous β-galactosidase-expressing organs is superior over the use of X-Gal/FeCN at slightly basic pH conditions.
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