Search results for "acrylamide"
showing 10 items of 485 documents
Applicability of the log MM - √D relationship to linear polyacrylamide gradient gel electrophoresis under a wide range of experimental conditions
1982
Recently we reported about a linear correlation between the logarithm of the size of native proteins (log mol mass or log Stokes' radius) and the square root of their migration distance (- √D) in linear polyacrylamide (PAA)-gradient gels (G. M. Rothe and H. Purkhanbaba, Electrophoresis 1982, 3, 33–42). The linearity between log MM and √D is not subject to time using homogeneous buffers in electrophoresis, no matter how the constants of the corresponding regression lines, slope and intercept change as a function of time. The realiability of this correlction has been re-examined with 0.7 mm thin gel plates and extending the time of electrophoresis under non-denaturating conditions from 2 to 9…
Efficient Extraction of Olive Pulp and Stone Proteins by using an Enzyme-Assisted Method
2014
An efficient protein extraction protocol for proteins from olive pulp and stone by using enzymes was developed. For this purpose, different parameters that affect the extraction process, such as enzyme type and content, pH, and extraction temperature and time, were tested. The influence of these factors on protein recovery was examined using the standard Bradford assay, while the extracted proteins were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The best extraction conditions were achieved at pH 7.0 and 5% (v/v) Palatase® 20000 L (lipase) for pulp and Lecitase® Ultra (phospholipase) for stone proteins. The optimal extraction temperature and time w…
Computer-aided calculation of the molecular size of nondenatured proteins in pore-gradient gel electrophoresis
1991
A computer program written in Turbo C is described, which uses the two-step mathematical procedure published recently (Rothe, G. M., Electrophoresis 1988, 9, 307-316) to evaluate the molecular mass, Stokes' radius, spherical radius, and frictional coefficient of nondenatured proteins. The program runs on any IBM-PC or 100% compatible IBM-PC, provided the disk operating system MS-DOS or PC-DOS 3.0 or later has been installed. Functions that are permanently in use are accessible by menu. Storage and loading of data from disk and help instructions can be called by use of function keys. The program provides several tables into which inserted and calculated data is automatically integrated. Each…
Sinusoidal endothelial cells from guinea pig liver synthesize and secrete cellular fibronectin in vitro.
1987
Endothelial liver cells were obtained from guinea pig by enzymatic digestion and centrifugal elutriation. Cells were cultured on gelatin and fibronectin pretreated culture vessels. Endothelial cells were characterized by phase-contrast microscopy, electron microscopy and the presence of Factor VIII-related antigen. Fibronectin secretion was determined in cell-free supernatants by a sensitive and specific ELISA and localized on fixed cultured cells by immunofluorescence. [35S]Methionine endogeneously labeled fibronectin was immunoprecipitated from supernatants and cellular lysates and displayed on sodium dodecyl sulfate polyacrylamide slab gel electrophoresis. After attachment to the culture…
Studies on Holothuriapolii (echinodermata) coelomocyte lysate II. Isolation of coelomocyte hemolysins
1988
The lytic activity of the Holothuria polii coelomocyte lysate resides in two electrophoretically distinct hemolysins identified as He1 and He2. He1 represents the calcium dependent, heat-labile component whereas He2 is calcium independent and heat-stable. The two hemolysins share serological identity. Both hemolysins appear as single protein molecules of 80KDa molecular weight by SDS-PAGE and transblotting analysis under non-reducing conditions. However under reducing conditions, they are doublets of 76 and 80KDa molecular weight. The hypothesis that the two hemolysins could be isoforms is discussed.
Use of voltage gradient gel electrophoresis in apoptotic DNA analysis
2000
In this paper the use of voltage gradient gel electrophoresis (VGGE) in the electrophoretic analysis of apoptotic DNAs is described. The peculiarity of VGGE fractionation in enhancing DNA bands in the gel by reducing their thickness was used to obtain a rapid, more selective and higher-quality electrophoretic fractionation of apoptotic DNA with respect to conventional electrophoresis. The use of VGGE fractionations also allowed a reduced amount of DNA to be used to detect a characteristic apoptotic DNA ladder pattern, in a lower agarose gel concentration, with respect to conventional electrophoretic fractionation
Isolation and partial characterization of a cytochrome-o complex from chromatophores of the photosynthetic bacterium Rhodospirillum rubrum FR1.
1989
A cytochrome-o complex was isolated from chromatophores of photoheterotrophically grown Rhodospirillum rubrum FR1. The enzyme was extracted with the non-denaturating detergent taurodeoxycholate and subsequently purified by sucrose-density-gradient centrifugation and gel-permeation HPLC. The complex contains two types of cytochromes, one of them cytochrome o, and two copper atoms. It catalyzes the reduction of molecular oxygen, when N,N,N',N'-tetramethyl-p-phenylenediamine or ubiquinol 10 are offered as electron donors. The oxidase activity is inhibited by cyanide, carbon monoxide and 2-heptyl-2-hydroxyquinoline N-oxide. The molecular mass of the protein is 136 +/- 15 kDa. The subunit analys…
Histone variants from pea (Pisum sativum): Their differential presence in fractions obtained by DNase I digestion of nuclei
1990
The variants of the core histones of Pisum sativum L. cv. Lincoln have been resolved by two dimensional polyacrylamide gel electrophoresis. Acetic acid, 8 M urea, 7.2 mM Triton X-100 was used in the first dimension. The second dimension was run in the presence of either anionic (sodium dodecylsulphate) or cationic (cetyltrimethyl-aminonium bromide) detergents. Four putative variants were found for the H2B histone class, 4 for H3 and 3 for H2A. Peptide mapping with (Staphylococcus aureus V8 protease was used, together with other criteria, to characterize the variants. The pattern of histone variants is not organ specific and, in an attempt to determine whether the diversity of histone varian…
Determination of molecular mass, Stokes' radius, frictional coefficient and isomer-type of non-denatured proteins by time-dependent pore gradient gel…
1988
Molecular mass, Stoke's radius, frictional coefficient and isomer-type of non-denatured proteins can be obtained by time-dependent gradient gel electrophoresis by evaluating the resulting data using a two-step mathematical procedure. Provided a histochemical staining procedure is available to locate the position of an enzyme in the gel, crude cell extracts can be used for estimating their molecular size properties. The computation of molecular properties of non-denatured proteins is demonstrated for isozymes of aspartate aminotransferase (EC 2.6.1.1), peroxidase (EC 1.11.1.42) and glucose-6-phosphate dehydrogenase (EC 1.1.1.49) from current-year needles of spruce. The resulting data as well…
Fluorescence of soil humic acids and their fractions obtained by tandem size exclusion chromatography-polyacrylamide gel electrophoresis
2002
Humic acids (HAs) extracted from soils of different origin (chernozem, ferralsol and ranker) and their fractions (A, B and C+D) obtained by tandem size exclusion chromatography–polyacrylamide gel electrophoresis were investigated by steady-state fluorescence spectroscopy in the emission mode. Independently of HA source, high molecular size fractions A and B are shown to be weakly fluorescent. The main fluorophores, especially those emitting at long wavelength (around 500–510 nm), are contained in the polar and low molecular size fractions C+D. As indicated by the observed pH effect, aromatic structures bearing carboxylate and OH substituents may be involved in these longer wavelength emissi…