Publications (98) View all
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Article: Primary structure of osteocalcin from ovine bone
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ABSTRACT: Osteocalcin was isolated from ovine (sheep) bone and purified by chromatography. The primary structure was determined using a manual micro-DABITC/PITC double coupling method. Ovine osteocalcin contains two modified amino acids, 4-hydroxyproline and -carboxyglutamate. These residues were readily identified in the sequencing process and the data presented here extend the micro manual sequencing method to include identification of 4-hydroxyproline and -carboxyglutamate residues. Ovine osteocalcin consists of 49 amino acid residues and the sequence confirms the high degree of conservation previously observed between bovine and human osteocalcins.European Journal of Allergy and Clinical Immunology 01/2009; 24(3):297 - 302. · 1.30 Impact Factor -
Chapter: The Electrophoretic Elution of Proteins from Polyacrylamide Gels
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ABSTRACT: The analytical power of acrylamide gel electrophoresis is one of the keys of modern protein chemistry. It is not surprising, therefore, that many methods have been described for converting that analytical power into a preparative tool. None of the available methods are entirely satisfactory for general use since loss of resolution or low recovery is often involved. The method described here has given both high resolution and good recovery but suffers from the disadvantage of being relatively laborious (1,2). In addition, although the recovered proteins are good for peptide analysis or amino acid composition determination, we have found very low yields on Edman degradation of proteins eluted from gels (3).03/2008: pages 169-175; -
Chapter: Fluorography of Polyacrylamide Gels Containing Tritium
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ABSTRACT: Fluorography is the term used for the process of determining radioactivity in gels and other media by a combination of fluorescence and photography. Since most of the radiation of a low energy emitter will largely be absorbed by the gel, in the technique of fluorography a fluor (e.g., PPO) is infiltrated into the gel where it can absorb the radiation and re-emit light that will pass through the gel to the film. The resulting photographic image is analogous to an autoradiograph, but for low energy (β-emitting isotope like 3H, the sensitivity of fluorography is many times the sensitivity of autoradiography. The fluorograph may be used directly, as a qualitative picture of the radioactivity on the gel. It may also be used to locate radioactive bands or spots that can then be cut from the original gel for further analysis, or be scanned to give quantitative information about the distribution of radioactivity. Figure 1 shows an example of a gel that was stained with Coomassie blue and then fluorographed. Notice that there is no loss of resolution in the fluorography of thin gels of normal size.03/2008: pages 163-167; -
Chapter: Computer Analysis of Gel Scans
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ABSTRACT: Gels are frequently scanned either to obtain quantitative data from the patterns of stain or radioactivity or to facilitate comparisons between samples. Both these processes can be greatly enhanced by using a computer and I will describe the use of a desktop or microcomputer in this area. There are several programs available for scanning and analyzing two-dimensional gels and I will mention these briefly, in passing. These programs need extensive computer facilities and a full discussion of these systems is beyond the scope of this chapter (see ref. 1 and references therein). One-dimensional gel scans, usually individual tracks from a slab gel, are routinely used for estimating the quantities of specific proteins, or the amounts of radioactivity in specific proteins. I will describe an interactive integration program that provides very flexible procedures for determining the areas under peaks in the scan. A more automatic integration system can also be used and a number of such systems, designed for chromatography, are available. I will describe a peak picking routine that could be used to develop an automatic integration program if commercial software is not available.02/2008: pages 127-139; -
Chapter: The Isolation of Satellite DNA by Density Gradient Centrifugation
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ABSTRACT: The term satellite DNA is used for a DNA component that gives a sharp band in a density gradient and can be resolved from the broader main band of DNA in the gradient. The usual gradient material is CsCl in aqueous buffer and the Cs+ ions form a density gradient in a centrifugal field. DNA in the solution sediments to its isopycnic point. The density of DNA is a function of base composition and sequence and so a homogeneous or highly repeated DNA sequence will form a sharp band in CsCl density gradients at a characteristic density. The resolution of this procedure may be enhanced or modified by binding ligands to the DNA. For example, netropsin binds specifically to A + T-rich regions of DNA and reduces their density (1,2). Another useful ligand is Ag+, which must then be centrifuged in Cs2SO4 gradients to avoid precipitation of AgCl (3). Pharmacia has recently introduced CsCF3COO as a gradient material.02/2008: pages 21-29;
