Double fluorescent in situ hybridization to zebrafish embryos.
AuthorsJowett T, Yan YL
JournalTrends Genet
PubMed ID8909127
Double in situ hybridization on mouse embryos for detection of overlapping regions of gene expression.
AuthorsBueno D, Skinner J, Abud H, Heath JK
JournalTrends Genet
PubMed ID8909126
Photobleaching of arterial autofluorescence for immunofluorescence applications.
AuthorsKingsley K, Carroll K, Huff JL, Plopper GE
JournalBiotechniques
PubMed ID11314262
Immunohistochemical localization of low-level antigens in the arterial vasculature is complicated by the presence of complex molecules such as collagen, elastin, cholesterol, and fluorescent lipids that exhibit autofluorescence over a wide spectrum of wavelengths. UV irradiation of arterial vasculature has remained ineffective in preparing samples for immunofluorescent staining because of ... More
Laminin-5 induces osteogenic gene expression in human mesenchymal stem cells through an ERK-dependent pathway.
AuthorsKlees RF, Salasznyk RM, Kingsley K, Williams WA, Boskey A, Plopper GE
JournalMol Biol Cell
PubMed ID15574877
The laminin family of proteins is critical for managing a variety of cellular activities including migration, adhesion, and differentiation. In bone, the roles of laminins in controlling osteogenic differentiation of human mesenchymal stem cells (hMSC) are unknown. We report here that laminin-5 is found in bone and expressed by hMSC. ... More
Use of a new fluorogenic phosphatase substrate in immunohistochemical applications.
AuthorsLarison KD, BreMiller R, Wells KS, Clements I, Haugland RP
JournalJ Histochem Cytochem
PubMed ID7822768
We used the phosphatase substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6- chloro-4-[3H]-quinazolinone, with standard alkaline phosphatase-mediated immunohistochemical techniques, to visualize a number of antibodies that bind to adult zebrafish retinal tissue. This compound, known as the ELF (enzyme-labeled-fluorescence) phosphatase substrate, produces a precipitate that fluoresces at approximately 500-580 nm (bright yellow-green). We show that the ... More
2-(2'-phosphoryloxyphenyl)-4(3H)-quinazolinone derivatives as fluorogenic precipitating substrates of phosphatases.
AuthorsHuang Z, Terpetschnig E, You W, Haugland RP
JournalAnal Biochem
PubMed ID1336935
Characterization of 2-(2'-phosphoryloxyphenyl)-4(3H)-quinazolinone (PPQ) derivatives as fluorogenic precipitating substrates of phosphatases is reported in this work. Soluble and colorless PPQ derivatives can be specifically hydrolyzed by acid and alkaline phosphatases into insoluble products, 2-(2'-hydroxyphenyl)-4(3H)-quinazolinone (HPQ) derivatives which appear as fluorescent precipitates in water. The fluorescence and precipitation of HPQ depend ... More
Functional expression and characterization of the wild-type mammalian renal cortex sodium/phosphate cotransporter and an 215R mutant in Saccharomyces cerevisiae.
AuthorsBernhardt F, Schoner W, Schroeder B, Breves G, Scheiner-Bobis G
JournalBiochemistry
PubMed ID10521262
The wild-type and an R215E mutant of the rat renal cortex sodium/phosphate cotransporter type 2 (NaPi-2) were functionally expressed in the yeast Saccharomyces cerevisiae strain MB192, a cell line lacking the high-affinity endogenous H+/P(i) cotransporter. The expression of the mRNA molecules and corresponding proteins was confirmed by Northern and Western ... More
Detection of endogenous and antibody-conjugated alkaline phosphatase with ELF-97 phosphate in multicolor flow cytometry applications.
AuthorsTelford W, Cox W, Singer V
JournalCytometry
PubMed ID11169576
BACKGROUND: The fluorogenic alkaline phosphatase (AP) substrate 2-(5'-chloro-2'-phosphoryloxyphenyl)-6-chloro-4-(3H)-quinazolinone (ELF(R)-97 phosphate, for Enzyme-Labeled Fluorescence) has been used primarily in microscope-based imaging applications to detect endogenous AP activity, antigens and various ligands in cells and tissues, and nucleic acid hybridization. In a previous study, we demonstrated the applicability of ELF-97 phosphate for ... More
Transmembrane topology of alpha- and beta-subunits of Na+,K+-ATPase derived from beta-galactosidase fusion proteins expressed in yeast.
AuthorsFiedler B, Scheiner-Bobis G
JournalJ Biol Chem
PubMed ID8910592
Various models of the transmembrane topology of the Na+,K+-ATPase predict either 8 or 10 membrane spans for the alpha-subunit and one to three membrane spans for the beta-subunit. Structure/function analysis, however, requires precise knowledge about the folding of enzymes. Therefore, the intention of this work was to establish a transmembrane ... More