» Articles » PMID: 28475610

An Improved Method with High Sensitivity and Low Background in Detecting Low β-galactosidase Expression in Mouse Embryos

Overview
Journal PLoS One
Date 2017 May 6
PMID 28475610
Citations 3
Authors
Affiliations
Soon will be listed here.
Abstract

LacZ is widely used as a reporter in studies of gene expression patterns. β-galactosidase, the product of LacZ gene, is usually detected by X-gal/FeCN staining. In X-gal/FeCN staining, β-galactosidase catalyzes X-gal to produce blue precipitates, which indicate the expression patterns of the gene of interest. A newer LacZ detection method using S-gal/TNBT is more sensitive but plagued by high background. Here, we describe an improved procedure that combines advantageous steps from the two methods. By comparing with X-gal/FeCN and S-gal/TNBT methods in detecting the expression patterns of miR-322/503 and miR-451 at a series of developmental stages, the improved method showed higher sensitivity and lower background. Thus, the improved method could be an alternative way of β-galactosidase staining in low gene expression situations.

Citing Articles

Evidence for Fgf and Wnt regulation of during limb development via two limb-specific -associated -regulatory modules.

Britton J, Somogyi-Leatigaga A, Watson B, Haro E, Mulder C, Kennedy K Front Cell Dev Biol. 2025; 13:1552716.

PMID: 40052149 PMC: 11882541. DOI: 10.3389/fcell.2025.1552716.


An Improved Staining Method for Low Signal Reporter Detection in Mouse Embryos.

Shen X, Xu F, Wu S, Li M, Zhang J, Liang R Bio Protoc. 2021; 9(5):e3180.

PMID: 33654983 PMC: 7854205. DOI: 10.21769/BioProtoc.3180.


Identifying vitamin A signaling by visualizing gene and protein activity, and by quantification of vitamin A metabolites.

Shannon S, Yu J, Defnet A, Bongfeldt D, Moise A, Kane M Methods Enzymol. 2020; 637:367-418.

PMID: 32359653 PMC: 7565286. DOI: 10.1016/bs.mie.2020.03.011.


Tracing Gene Expression Through Detection of β-galactosidase Activity in Whole Mouse Embryos.

Blanco M, Learte A, Marchena M, Munoz-Saez E, Cid M, Rodriguez-Martin I J Vis Exp. 2018; (136).

PMID: 30010638 PMC: 6101999. DOI: 10.3791/57785.

References
1.
Soibam B, Benham A, Kim J, Weng K, Yang L, Xu X . Genome-Wide Identification of MESP1 Targets Demonstrates Primary Regulation Over Mesendoderm Gene Activity. Stem Cells. 2015; 33(11):3254-65. DOI: 10.1002/stem.2111. View

2.
Shen X, Soibam B, Benham A, Xu X, Chopra M, Peng X . miR-322/-503 cluster is expressed in the earliest cardiac progenitor cells and drives cardiomyocyte specification. Proc Natl Acad Sci U S A. 2016; 113(34):9551-6. PMC: 5003281. DOI: 10.1073/pnas.1608256113. View

3.
Stefanescu E, Dorneanu M, Danila C, Aprotosoaie C, Grosu G . Tetrazolium salts and metal complexes of some formazans. Rev Med Chir Soc Med Nat Iasi. 2000; 103(1-2):182-5. View

4.
Cotson S, Holt S . Studies in enzyme cytochemistry. IV. Kinetics of aerial oxidation of indoxyl and some of its halogen derivatives. Proc R Soc Lond B Biol Sci. 1958; 148(933):506-19. DOI: 10.1098/rspb.1958.0042. View

5.
Peng X, Shen X, Chen X, Liang R, Azares A, Liu Y . Celf1 regulates cell cycle and is partially responsible for defective myoblast differentiation in myotonic dystrophy RNA toxicity. Biochim Biophys Acta. 2015; 1852(7):1490-7. DOI: 10.1016/j.bbadis.2015.04.010. View