Protein microchips: use for immunoassay and enzymatic reactions.
AuthorsArenkov P, Kukhtin A, Gemmell A, Voloshchuk S, Chupeeva V, Mirzabekov A
JournalAnal Biochem
PubMed ID10660453
'Different proteins such as antibodies, antigens, and enzymes were immobilized within the 100 x 100 x 20-microm gel pads of protein microchips. A modified polyacrylamide gel has been developed to accommodate proteins of a size up to 400,000 daltons. Electrophoresis in the microchip reaction chamber speeded up antigen-antibody interactions within ... More
Optimization of heterologous gene expression for in vitro evolution.
AuthorsMatsumura I, Olsen MJ, Ellington AD
JournalBiotechniques
PubMed ID11252780
Rainbow's end: the quest for multiplexed fluorescence quantitative analysis in proteomics.
AuthorsPatton WF, Beechem JM
JournalCurr Opin Chem Biol
PubMed ID11827825
During the past two years, the performance of fluorescence-based protein detection methods has demonstrably eclipsed conventional technologies such as colloidal Coomassie Blue and silver staining with respect to detection sensitivity, quantitative accuracy and compatibility with modern protein identification and characterization procedures. At this point, fluorescence-based methods are poised to offer ... More
A fluorogenic substrate for beta-glucuronidase: applications in fluorometric, polyacrylamide gel and histochemical assays.
AuthorsZhou M, Upson RH, Diwu Z, Haugland RP
JournalJ Biochem Biophys Methods
PubMed ID9029263
We have developed a fluorogenic substrate, ELF-97 beta-D-glucuronide, that provides significant advantages over existing substrates in detecting beta-glucuronidase activity. ELF-97 beta-D-glucuronide allows the detection of enzymatic activity in situ, yielding a hydrolytic product that exhibits maximal fluorescence within the physiological pH range. This substrate yields a hydrolytic product that demonstrates ... More
Simultaneous, two-color fluorescence detection of total protein profiles and beta-glucuronidase activity in polyacrylamide gel.
AuthorsKemper C, Steinberg TH, Jones L, Patton WF
JournalElectrophoresis
PubMed ID11332766
A dichromatic method for measuring the specific activity of beta-glucuronidase from complex cell homogenates or partially purified protein fractions is presented. Dual fluorescence is achieved by using the green emitting fluorogenic substrate ELF 97 beta-D-glucuronide to detect beta-glucuronidase activity, followed by the red emitting SYPRO Ruby protein gel stain or ... More
Klotho is a novel beta-glucuronidase capable of hydrolyzing steroid beta-glucuronides.
AuthorsTohyama O, Imura A, Iwano A, Freund JN, Henrissat B, Fujimori T, Nabeshima Y
JournalJ Biol Chem
PubMed ID14701853
klotho mutant mice provide a unique model to analyze mechanisms of aging because their phenotypes resemble those of human aging-associated disorders. The klotho gene encodes Klotho, a type I membrane protein that shares sequence similarity with members of the glycosidase family 1. Because Klotho lacks the glutamic acid residues that ... More
Fluorescence detection of proteins in sodium dodecyl sulfate-polyacrylamide gels using environmentally benign, nonfixative, saline solution.
SYPRO Tangerine stain is an environmentally benign alternative to conventional protein stains that does not require solvents such as methanol or acetic acid for effective protein visualization. Instead, proteins can be stained in a wide range of buffers, including phosphate-buffered saline or simply 150 mM NaCl using an easy, one-step ... More