Electrophoresis on gels

Biotechnology is being used in the diagnosis of disease. For example, samples of DNA are treated with restriction endonucleases and then subjected to electrophoresis on gels. Using the Southern blot technique, the x restriction fragments on the gel are hybridized with radioactive cDNA... 

The molecular weight of a protein and its net charge may be determined by performing electrophoresis on gels with constant %C, by varying %T from gel to gel. After separation, the migration of the protein is measured relative to a tracking dye, and the Rf value is calculated ... 

The original technique developed by Weber and Osbron [94] exists today in two variations with regard to the spatial arrangement in the form of disc (discontinuous) electrophoresis on gel rods, or in the form of slab gel separation. 

The analysis of human body fluids and tissue extracts by electrophoresis and clinical applications of the technique is one of the most rapidly developing areas of biological research. Whilst analysis of serum proteins by one-dimensional electrophoresis on gel-media or even cellulose acetate still continues to produce... 

Here A stands for co-electrophoresis on gels with marker protein B, co-chromatography with marker protein C, specific tryptic peptides identified and D, precipitation by specific antibody. 

The molecular mass of the monomer of GLO is about 50,000 daltons (Kiuchi et al., 1982 Nishikimi et al., 1976), as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. On gel filtration the native enzyme behaves as an aggregate with a molecular mass of about 500,000 daltons for rat and goat enzymes (Nishikimi et al., 1976) and of about 400,000 daltons for chicken enzyme (Kiuchi et al., 1982). 

A number of different approaches have been taken to describing transport in porous media. The objective here is not to review all approaches, but to present a framework for comparison of various approaches in order to highlight those of particular interest for analysis of diffusion and electrophoresis in gels and other nanoporous materials. General reviews on the fundamental aspects of experiments and theory of diffusion in porous media are given... 

CK isoenzymes have been separated by electrophoresis on cellulose acetate, agar gel, agarose and polyacrylamide gel. 

Figure 3. SDS-PAGE and in situ pectinase activity on pectin and polygalacturonic acid-agarose overlays of culture filtrates of Aspergillus niger N-402 (upper panel) and Aspergillus FP-180 (lower panel) at 2.5, 3.5, 5.5 and 6.5 pHi (Lanes a, b, c, and d, respectively). Electrophoresis on 10% acrylamide slab gel (14 X 8 cm) in the presence of SDS was according to Laemmli (6), run at 30 mA constant current for 2 hours. Crude cell-free samples were concentrated by lyophilization, dialyzed, boiled with sample buffer by 60 sec. and applied to each well. Polyacrylamide gel and overlays were incubated overnight with 0.17 acetate buffer at room temperature.

By means of gel electrophoresis on cross-linked, hydrolyzed starch,99 with simultaneous checking for proteins, lipids, and pectinesterase activity, it was found, however, that the product isolated after the separation on CM-Sephadex C-50 constitutes but one of five multiple forms of tomato pectinesterase, and is the one present in preponderant proportion98 (see Fig. 4). The accompanying lipid and sugar components were separated from this pectinesterase form in the course of the purification procedure. After analysis of the hydro-lyzate of the final product for fatty acids, as well as for carbohydrate components, it was possible to exclude the possibility of a lipoprotein,30 as well as glycoprotein,100 character of this form of tomato pectinesterase. 

Preparative electrophoresis on Sephadex G-25 (Ref. 168) or double isoelectric focusing,208 preceded by chromatography on Sephadex G-75, CM-cellulose, and calcium phosphate, was used for the isolation of endo-D-galacturonanase from the filtrate of a Verticillium albo-atrum culture. The homogeneity was confirmed in both cases by electrophoresis on poly(acrylamide) gel. The molecular weight of the enzyme was close to the values found for Aspergillus endo-D-galacturonanases. 

Figure 7.10. IL-l/J expression by GM-CSF-stimulated neutrophils. Human neutrophils were incubated in the absence (-) and presence (+) of 50 U/ml GM-CSF, and at time intervals total RNA was extracted. After electrophoresis on formamide-containing gels, IL-1/3 mRNA levels were probed. Source Experiment of Julie Quayle.

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