The lacZ gene product, β-galactosidase, has found extensive use as a reporter of gene expression. Many methods for detecting lacZ expression in transformed cells are limited because they can require harsh conditions that affect viability of the cells after detection. To overcome this problem, lipophilic substrates for detection of lacZ expression in living cells, such as C₁₂FDG have been developed that can be used under standard culture or physiological conditions. After incubation with these fluorogenic substrates, cultured lacZ-positive mammalian cells appear morphologically normal, continue to divide, and retain the lipidic fluorescent product for long periods. Because the product is so well retained, fluorescence intensity can be quantitatively related to the level of gene expression. Because of its lipophilic nature, C₁₂FDG has also found extensive use in the analysis of lacZ and SA-bGal activity in tissue sections, tissue biopsies and in vivo where is it well retained for analysis.
C₁₂FDG; C12-FDG; C12 FDG; C12FDG
MeOH, H₂O (sl.)
-20C, Protect From Light, Desiccated
References and Citations
Cooper MA, Zhou R. (2013) "β-Galactosidase staining of lacZ fusion proteins in whole tissue preparations." Methods Mol Biol. 1018:189-197.
Noppe G, Dekker P, de Koning-Treurniet C, Blom J, van Heemst D, Dirks RW, Tanke HJ, Westendorp RG, Maier AB. (2009) "Rapid flow cytometric method for measuring senescence associated beta-galactosidase activity in human fibroblasts." Cytometry A. 75(11):910-916.
Debacq-Chainiaux F, Erusalimsky JD, Campisi J, Toussaint O. (2009) "Protocols to detect senescence-associated beta-galactosidase (SA-betagal) activity, a biomarker of senescent cells in culture and in vivo." Nat Protoc 4:1798-1806.
Okamoto Y, Monjushiro H, Fukumoto T, Watarai H. (2006) "Measurement of hydrolysis kinetics of galactose-substituted fluorescein by beta-galactosidase at the toluene-water interface by spinning microtube fluorometry." Anal Bioanal Chem 385:1430-1438.
Plovins A, Alvarez AM, Ibañez M, Molina M, Nombela C. (1994) "Use of fluorescein-di-beta-D-galactopyranoside (FDG) and C12-FDG as substrates for beta-galactosidase detection by flow cytometry in animal, bacterial, and yeast cells." Appl Environ Microbiol. 60(12):4638-4641.
Zhang YZ, Naleway JJ, Larison KD, Huang ZJ, Haugland RP. (1991) "Detecting lacZ gene expression in living cells with new lipophilic, fluorogenic beta-galactosidase substrates." FASEB J. 5:3108-3113.
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