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In the diagnosis of alcohol abuse transferrin (Tf) allelic D variants generate false-positive test results for carbohydrate-deficient transferrin (CDT) as assessed by their electrophoretic migration patterns. The predominant Tf C1 allele encodes a protein for which the most prevalent isoform has a pI of 5.4, i.e., four sialic acids and two bound ion molecules. Carriers of allele D encode Tfs wi...
Biochemical study of thymus leukemia antigen (TL) from thymocytes of various Tla genotypes and from leukemia cells revealed features that, given present evidence, are peculiar to TL among class I products of the H-2:Qa:Tla region of chromosome 17. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of TL from thymocytes of all TL+ mouse strains, precipitated by anti-TL antiseru...
Two-dimensional gel electrophoresis (2-DE) is able to separate hundreds to thousands of proteins or polypeptides by coupling IsoElectric Focusing (IEF) in first dimension and Sodium Dodecyl Sulphate PolyAcrylamide-Gel Electrophoresis (SDS-PAGE) in second dimension. This particular configuration is called classical 2-DE: IEF separates proteins in function of their isoelectric point (pI) and SDS-...
Isoelectric focusing (IEF) profiles of pectate lyases (PLs) produced by five different groups of soft rot bacteria were analyzed by using the combined techniques of thin-layer polyacrylamide gel IEF and agarose-pectate overlay activity staining. Four strains of soft rot Erwinia spp. produced three or more PL isozymes. All of eight Pseudomonas viridiflava strains examined produced one single PL ...
Artifacts in two-dimensional electrophoresis (2-DE) caused by the presence of salts in isoelectric focusing (IEF) have been previously described as a result of increasing conductivity and inducing electroosmosis. However, electrolysis induced by the presence of salts should not be disregarded. In this study, electrolytic reduction-oxidation reaction (redox) was found to be enhanced in the prese...
Biochemical and serological studies were performed on more than 400 anti- phosphocholine (PC) hybridoma proteins (HP) derived from six strains of mice; 26 of these HP were examined in detail. All HP possessed specificity for PC, and all those tested contained an H-chain idiotypic determinant, V(H)-PC, which is shared by PC-binding myeloma proteins (BMP) and anti-PC antibodies. Among the HP, thr...
Primary biliary cirrhosis (PBC) is one of the few nonalcohol-induced liver pathologies that causes false positives in assays of carbohydrate-deficient transferrin (CDT) for diagnosing alcohol abuse. CDT was quantified by isoelectric focusing-immunoblotting-laser densitometry (IEF-IB-LD) analysis of serum from 117 women: 57 PBC patients, 20 alcohol abusers, and 40 healthy donors. Only 5% (3 of 5...
Two dimensional gel electrophoresis (2-DE), isoelectric focusing (IEF) and agarose gel electrophoresis (AGE) were used to examine cerebrospinal fluid (CSF) and sera from 22 patients with confirmed multiple sclerosis, 11 patients with probable multiple sclerosis and 20 control patients with non-inflammatory neurological diseases of the central nervous system (CNS). All of the 22 patients with co...
Complex protein mixtures have traditionally been separated by 2-DE. Görg introduced IPGs as the first dimension of protein separation. In recent years, MS-based proteomics has increasingly become the method of choice for identifying and quantifying large number of proteins. In that technology, to decrease analyte complexity, proteins are often separated by 1-D SDS-gel electrophoresis before onl...
The identification of 114 integral membrane proteins from Halobacterium salinarum was achieved using liquid chromatography/tandem mass spectrometric (LC/MS/MS) techniques, representing 20% of the predicted alpha-helical transmembrane proteins of the genome. For this experiment, a membrane preparation with only minor contamination by soluble proteins was prepared. From this membrane preparation ...
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