ISSN:0173-0835    

DOI:10.1002/elps.1150020102

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractMembrane proteins and cytosol proteins from organs (liver, brain) of inbred mice were separated by two‐dimensional electrophoresis (2DE). The 2DE technique revealed high resolution of complex protein solutions bystainingthe proteins. The 2DE patterns of membrane proteins showed about 660 protein spots (polypeptides) for both liver and brain. The protein patterns of the cytosols revealed about 800 polypeptides for the liver and about 680 for the brain. The membrane proteins and cytosol proteins of the 2DE patterns represented two protein populations specific for these two cell components. They matched in 7 % (liver) or 23 % (brain) of the total number of membrane protein spots. The matched spots represented dissociable membrane proteins rather than contamination of the isolated membrane fractions by cytosol proteins. Protein patterns of the same cell fraction but from the two different organs investigated contained more than 50 % organ‐unspecific polypeptides.Several conditions of our 2DE standard procedure were reinvestigated to test the usefulness for our technique of some methodical steps employed in 2DE by other authors. We found that many steps often used in sample preparations impaired rather than improved the 2DE pattern. In particular, addition of SDS to the protein sample resulted in a considerable loss of proteins during isoelectric foucing. The 2DE itself could be simplified by omitting several steps such as prefocusing, equilibration of focusing gels, forming gel gradients, autoradiography. This has some implications for the development of 2DE to a routine met

ISSN:0173-0835    

DOI:10.1002/elps.1150020103

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractAnalytical capillary isotachophoresis has been applied as an analytical tool for the assay of the microsomal enzyme UDP‐glucuronyltransferase, which is an important system for the detoxification of drugs and many endogenous and exogenous toxins. The substrate donor molecule UDP‐glucuronic acid, various products of hydrolytic cleavage, and the products of glucuronidation β‐glucuronides, can be assayed in a single run. Furthermore, the glucuronidation of mixtures of acceptors can be monitored simultaneously, since the various β‐glucuronides are separated from one another. This method offers a more comprehensive assay of glucuronidation reactions than has been possible using previously published p

ISSN:0173-0835    

DOI:10.1002/elps.1150020104

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractAn asymptotic solution to the partial differential equation governing solute behavior in the continuous flow electrophoresis device is combined with solutions for the convective, electrophoretic and electroosmotic velocities in the chamber under nonisothermal operating conditions. The results illustrate the effect of temperature gradients on the net dispersion of solute in continuous flow electrophoresis. Three modes of operation are tested for this chamber: the convective velocity is in the same direction as gravity, the convective velocity is in the direction opposite to gravity and the effect of gravity is negligible.

ISSN:0173-0835    

DOI:10.1002/elps.1150020105

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractA method for continuous preparative electrophoresis is presented. It is based on the principle that a supporting medium,e.g.a polyacrylamide gel, is moved continuously relative to an electrical field. At one fixed point, the sample is applied to the gel. The charged particles migrate with different velocities in the supporting medium. Simultaneously, they are dragged along with the supporting medium. This procedure leads to a two‐dimensional separation of charged particles, which is necessary for a continuous operation of an electrophoresis apparatus. Three different designs of apparatuses, using this method, are presented in this paper. The operation of one construction was tested by separating three protein mixture

ISSN:0173-0835    

DOI:10.1002/elps.1150020106

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractMouse splenic lymphocytes were examined for electrophoretic mobility by an automatic cell electrophoresis. The whole population of lymphocytes showed a bimodal histogram of electrophoretic mobility. In contrast, adherent cells to a nylon wool column (B cell‐rich) and nonadherent cells treated with anti‐mouse IgG and complement (T cell‐rich) gave a unimodal histogram. A linear relationship existed between the electrophoretic mobility of mixture of T and B cells and their ratio detected by the method of immunofluorescence, suggesting that the ratio of T to B cells can easily be determined from the electrophoretic mobility of lympho

ISSN:0173-0835    

DOI:10.1002/elps.1150020107

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractA simple Technique for horizontal gradient polyacrylamide gel electrophoresis (PAGE) in discontinuous buffer systems is described, in which one‐dimensional, two‐dimensional and double one‐dimensional separation patterns can be obtained. The advantages of this technique are: (i) that samples can be applied by overlaying on a variable broad area of the concentrating gel, (ii) that the complete separating gel and the contact with the concentrating gel remain sandwiched between glass plates from the moment of gel polymerization till the end of electrophoretic separation, (iii) that many gels are poured from one gel solution, and (iv) that the gels can be stored, due to an identical buffer composition in the concentrating and separatin

ISSN:0173-0835    

DOI:10.1002/elps.1150020108

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractBovine Submaxillary mucin (BSM) and human submandibular mucin (HSM) were electrophoresed in a system in which the mucins were incorporated into an agarose sample gel and separation was carried out in a 5 % polyacrylamide gel. BSM was resolved into two protein‐ and carbohydrate‐containing bands, and a number of non‐mucin proteins. When the electrophoresis was carried out at 80°C, the mobilities and resolution of the bands were increased. The entry of HSM into the running gel was also increased at the elevated temperature and the samples were separated into at least four protein‐ and carbohydrate‐stainable bands. Protein bound sialic acid remained unchanged unter these conditions and aggregating activity for cells of a strain of oral streptococcus could be recovered from the gels after the electrophoresis. The facilitation of the electrophoresis of human mucus glycoprotein's at high temperature may reflect the dissociation of macromolecular aggregates of the mucins under these conditions. A number of purified, non‐mucin proteins also moved as discrete bands at 80degreeC. It is suggested that electrophoresis at elevated temperatures should be considered generally for the analysis of multimolecular systems that are of biologica

ISSN:0173-0835    

DOI:10.1002/elps.1150020109

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

摘要:

AbstractA simple procedure is described for counting labeled protein bands after polyacrylamide slab gel electsrophoresis. Proteins labeled with radioisotopes are separated on conventional polyacrylamide slab gels. After fiction and staining, the slab gels are dried onto filter papers under vacuum. The labeled protein patterns are recorded by autoradiography or fluorography. The sample tracks are separated from each other by cutting the dried slab gel lengthwise. Each sample track is cut into predetermined sections by making parallel cuts with a film cutter. The sections are deposited into scintillation vials and digested with 50% hydrogen peroxide. The digested samples are counted in an aqueous scintillator. This procedure provides a direct quantitation of 3h and 14C‐labeled protein bands by reproducibility counting. The results also show that both high resolution and excellent reproducibility are obtainable by this relatively fast and inexpensive metho

ISSN:0173-0835    

DOI:10.1002/elps.1150020110

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150020111

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1981

数据来源: WILEY