共查询到20条相似文献,搜索用时 15 毫秒
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Wu L Wu J Zhang J Zhou Y Ren G Hu Y 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2008,867(1):62-68
A simple method was described for the purification of serum transferrin (Tf) from human plasma and porcine serum with relative high yield and purity. The properties including purity, integrity, immunoreactivity and the receptor-binding ability of the proteins were studied by several assays, comprising spectrometry, SDS-PAGE, HPLC, Western blotting, urea electrophoresis, mass spectrometry and cytometry. Analysis from all the different aspects manifested that the proteins were of high purity. The two kinds of Tfs appeared to be iron-saturated as confirmed by their absorbance spectra and urea-PAGE mobility. The specific spectra of absorption of the two Tfs were both at around 465 nm. The relative molecular weights of human Tf (hTf) and porcine Tf (pTf) were determined by SDS-PAGE and further identified by MAIDI-TOF mass spectrometry with a result of 79,707 and 79,258, respectively. Immunoblotting assay showed that pTf could react with the anti-human Tf monoclonal antibody with a less level compared to hTf. FACS assays of their binding activities to Tf receptor-positive cell (K562 cell line) indicated that pTf could be recognized by the hTf receptor and internalized into cells, with a slightly less efficacy than hTf. All special property studies demonstrated that pTf was similar to hTf in physical and chemical characteristics, which gave a hint that pTf could substitute for hTf in some kinds of researches, such as using hTf as a carrier in drug targeting system. 相似文献
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It is possible to solubilize selectively collagen from a collagen-elastin mixture by the use of a phenol:acetic acid:water solvent. This technique can be successfully applied to vascular tissue which has been salt-extracted and delipidated. As well as separating both of the major insoluble structural proteins from the one tissue sample, the method is very simple and inexpensive and requires minimal attention. 相似文献
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In this work we describe a simple two step separation procedure for the separation and purification of short DNA fragments. The first step involves precipitating the DNA using the cationic surfactant dodecyltrimethylammonium bromide. Dodecyltrimethylammonium bromide, unlike cetyltrimethylammonium bromide will not precipitate DNA before complexation is complete thus providing a high purity DNA. The second step involves dissolution of the DNA-dodecyltrimethylammonium complex in 75% ethanol, followed by precipitation of the Sodium-DNA salt, by titrating in a salt solution. This method is particularly suited to purification of short fragments as it does not require high salt concentrations in the ethanol precipitation step, which can be damaging for short DNA. The ability of dodecyltrimethylammonium bromide to remove ethidium bromide from intercalation sites on the DNA is also discussed 相似文献
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J Gniot-Szulzycka B Januszewska 《Folia histochemica et cytobiologica / Polish Academy of Sciences, Polish Histochemical and Cytochemical Society》1986,24(3):251-256
A method for rapid isolation of human placenta microsomes, which does not require facilities for ultracentrifugation was described. Such microsomes were compared with microsomes prepared by conventional ultracentrifugation technique. Both microsomal preparations were tested for protein, RNA and phospholipid content as well as for sulphohydrolase activities and glucose-6-phosphatase activity. The degree of contamination with other subcellular particles were tested as well. The data presented showed that microsomal proteins precipitated at pH 5.3 may be conveniently used for preparative separation of microsomal enzymes. 相似文献
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A simple one-step procedure for the separation of calpain I, calpain II and calpastatin. 总被引:4,自引:0,他引:4
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The soluble fraction from rabbit brain was adsorbed on a column of phenyl-Sepharose. By applying a linear gradient with decreasing salt concentration and increasing pH, it was possible to separate calpain I and calpain II from each other and from the endogenous inhibitor calpastatin. Both enzymes were capable of degrading endogenously labelled neuronal proteins, including slowly axonally transported soluble proteins and rapidly transported membrane-bound proteins, as well as casein. 相似文献
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Two human urinary kallikreins (fractions A-1 and A-2) were purified to apparently homogeneous forms. The two kallikreins were separated by affinity chromatography using Trasylol (aprotinin) covalently bound to Sepharose. The kallikreins were eluted with a pH gradient (pH 9.5-3.0). Fraction A-1 was eluted between pH 6.2 and 4.2 and fraction A-2 was eluted between pH 4.2 and 3.1. Final purification was obtained by chromatography on Sephacryl SS-200. Antibodies prepared against fraction A-2 were also reactive with fraction A-1; thus it may be possible to measure both by radioimmunoassay using the same antibody preparation. 相似文献
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Some physical-chemical properties of reduced-alkylated and sulphitolysed human serum transferrins and hen''s-egg conalbumin 总被引:3,自引:2,他引:1
Anatoly Bezkorovainy Dietmar Grohlich Claire M. Gerbeck 《The Biochemical journal》1968,110(4):765-770
1. Apparently all disulphide bridges of transferrin and conalbumin were broken by reduction-alkylation, whereas sulphitolysis resulted in incomplete cleavage of disulphide bonds. 2. The molecular weights of reduced-alkylated and sulphitolysed transferrin and reduced-alkylated conalbumin were identical with those of native proteins in a number of solvents, indicating that these proteins exist as single polypeptide chains. 3. Viscosity studies indicated that reduced-alkylated transferrin possesses a partially ordered structure in 0-4m-urea, assumes a random-coil configuration in 6m-urea with a molecular weight of 84000 and is partially aggregated in 8m-urea. 相似文献
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A new and simple colorimetric method for human serum lipase [EC 3.1.1.3] assay has been developed, using 2,3-dimercaptopropan-1-ol tributyroate as a substrate, 5,5'-dithiobis(2-nitro-benzoic acid) as a chromogenic reagent, phenylmethylsulfonyl fluoride as an inhibitor of serum esterases, and sodium dodecylsulfate as a lipase activator. The method requires only 50 micron1X2 of serum sample and a reaction time of less than 30 min. The method is reproducible and sensitive enough to measure low levels of lipase activity in normal and abnormal sera. The gel filtration of serum samples on a Sephadex G-200 column gave one peak of lipase activity, when measured by the present method, and the molecular weight of the enzyme was identical with that of lipase of human pancreatic origin, confirming the specificity of this new method for the serum lipase. 相似文献
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A simple procedure is described for the preparation of glucosamine and glucosaminitol, employing column chromatography on Sephadex G-10, using a borate-phosphate buffer as elutrient. 相似文献
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A method for the isolation of protein components by a combination of polyacrylamide gel electrophoresis and electro-extraction of slices of gel contained in cellophane dialysis tubing, using the H tube electrophoresis apparatus, is described. The method was applied to the purification of individual human serum glycoproteins. Although the glycoproteins have widely differing electrophoretic mobilities, they appeared to be antigenically interrelated judging from their complex "spurring" in Ouchterlony double gel diffusion tests. 相似文献
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A simple purification procedure for obtaining a high yield of electrophoretically and immunologically pure rat α-fetoprotein from amniotic fluid is described. Rat amniotic fluid is passed through an anti-rat albumin immunoabsorbent column to remove albumin. The albumin-free eluate is then chromatographed on DEAE-Sephacel to separate α-fetoprotein from transferrin and other minor protein contaminants. This two-step purification procedure results in a recovery of approximately 70% of the rat α-fetoprotein originally present in the amniotic fluid. 相似文献