摘要:

AbstractComputer simulation is used to analyze a system of two electrophoretic columns coupled by mixing the anolyte of one with the catholyte of the other. A mathematical model is presented which is used to predict the pH gradients formed by monovalent buffers in this system, when the currents in the columns are unequal. In the column with the higher current a pH gradient is created which increases from anode to cathode and is potentially useful for isoelectric focusing. The breadth of this gradient is dependent upon the ratio of the currents. The function of the second column is the compensation of buffer migration which occurs in the first column, thereby maintaining constant electrolyte composition. The effects of buffer pKs and mobilities are evaluated.

ISSN:0173-0835    

DOI:10.1002/elps.1150071102

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

年代:1986

数据来源: WILEY

摘要:

AbstractA new method for the apolipoprotein E (apo E) phenotyping directly from human serum has been developed. The method is based on isoelectric focusing of delipidated serum in vertical slab gel systems followed by blotting and immunostaining using polyclonal anti‐apo E antibodies as the first antibody. Apo E phenotyping with this method in 150 serum samples gave exactly the same results as obtained with the conventional method based on isoelectric focusing of delipidated very low density lipoproteins isolated by ultracentrifugation and detected by protein staining. Compared with the conventional method, the present method is less expensive, suitable for large‐scale screening purposese. g.diagnosis of type III hyperlipidemia and needs only a few microliters of serum. By application for screening of hyperlipidemic sera, we were able to detect two patients with an apo E‐1 variant and two others with “new” E‐3 and E‐4 mutants by the described methods. In addition apparent molecular weight variants of apo E could be detected by sodium dodecyl sulfate‐gel electrophoresis of sera followed by blotting and

ISSN:0173-0835    

DOI:10.1002/elps.1150071103

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

年代:1986

数据来源: WILEY

摘要:

AbstractUltrathin‐layer sodium dodecyl sulfate‐polyacrylamide gradient gel electrophoresis in combination with silver staining is a highly sensitive and rapid method for molecular size analysis of urinary proteins. First results are achieved after 4 h (electrophoretic run, 100 min, followed by Coomassie Brilliant Blue staining, 2 h). After additional silver staining, including a recycling step, the results are ready for evaluation within 7 h. Urine samples do not need to be concentrated. Within a single gel, 25–27 samples can be run side by side under identical conditions and thus compared without ambiguity. By coelectrophoresis of pure proteins and immunological analysis 13 urinary proteins could be identified and for 7 of them the detection limits were determined. Glomerular protein patterns in samples of urine with a protein concentration in the normal range are compared with the concentration of albumin and transferrin measured by a sensitive enzyme immunoassay. Typical renal and extrarenal protein patterns are

ISSN:0173-0835    

DOI:10.1002/elps.1150071104

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

年代:1986

数据来源: WILEY

摘要:

AbstractIsoelectric focusing in mixed carrier ampholyte‐immobilized pH gradients (CA‐IPG) is an effective way to separate proteins by charge. A method to prepare CA‐IPG in capillary tubes and the description of the equipment used are outlined. Two‐dimensional gel electrophoresis patterns of human serum and liver biopsies, with isoelectric focusing and CA‐IPG as the first dimension, are presented. The comparison with two‐dimensional gel electrophoresis patterns obtained by conventional carrier ampholyte pH gradient separation shows the excellent resolution and potential of thi

ISSN:0173-0835    

DOI:10.1002/elps.1150071105

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

年代:1986

数据来源: WILEY

摘要:

AbstractProgesterone therapy of androgenized rats results in the elimination of histological abnormalities of the uterus associated with constant unopposed oestrogenic stimulation. Subcellular fractions prepared from uteri of normal, androgenized and progesterone treated animals have been analysed by two‐dimensional gel electrophoresis. In all fractions, protein changes were observed as a result of androgenization and subsequent progesterone therapy. The identity of some of these proteins has been assigned following peptide mapping. Following progesterone therapy, the relative proportion of cytosolic tubulins, tropomyosins and D‐glucose‐6‐phosphate dehydrogenase were increased. In the abnormal uterus, a protein with similarity to the B subunit of creatine kinase was present in the mitochondrial fraction but could not be detected in the cytosol. Subsequent to progesterone therapy, this protein was no longer detectable in the mitochondrial fraction whilst two molecular forms of the B subunit of creatine kinase appeared in the cytosol. These hormonally‐sensitive changes in both structural and enzymatic proteins, which are ubiquitous to most cell types, are discussed in relation to morphological differentiation of the uterus and the molecular mechanisms underlying ce

ISSN:0173-0835    

DOI:10.1002/elps.1150071106

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

年代:1986

数据来源: WILEY

摘要:

AbstractWater soluble allergens of grass pollen (Dactylis glomerata) were purified by two successive steps of preparative isoelectric focusing in a flat bed of granulated gel and controlled by analytical isoelectric focusing in agarose gels. In order to separate components cofocusing in the same pH zone but having different electrophoretic mobilities, preparative zone electrophoresis in granulated gels was performed. The homogeneity of the fractions recovered was tested by isoelectric focusing and zone electrophoresis, followed by radioimmunodetection of the allergenic molecules on nitrocellulose prints. The major allergen ofDactylis glomeratapollen was purified with a yield of 7.5 mg from 10 g of pollen grains.

ISSN:0173-0835    

DOI:10.1002/elps.1150071107

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

年代:1986

数据来源: WILEY

摘要:

AbstractHair proteins have been successfully analysed by high resolution two‐dimensional electrophoresis by avoiding S‐carboxymethylation which has previously dictated the use of alternative two‐dimensional procedures. The high and low‐sulfur proteins are simultaneously resolved within the pIrange 3–9 and their electrophoretically determinedMrvalues are consistent with molecular weights obtained by physica

ISSN:0173-0835    

DOI:10.1002/elps.1150071108

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

年代:1986

数据来源: WILEY

摘要:

AbstractThe degradation kinetics of Immobiline pK9.3 (0.2 m in H2O) have been studied over a period of 2‐years in the frozen state. The apparent hydrolysis rate (derived from the shift of myosin spots in a two‐dimensional map) is 20% per year (or 10% if incorporation of acrylic acid, an hydrolysis product, is accounted for). Such reactions in frozen solutions are not uncommon (cf.the spontaneous mutarotation of glucose, Kiovsky, T. E. and Pincock, R. E.,J. Amer. Chem. Soc.1966,88, 4704–

ISSN:0173-0835    

DOI:10.1002/elps.1150071109

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

年代:1986

数据来源: WILEY

摘要:

AbstractThe human plasma transthyretin (TTR, prealbumin) variant TTR(Met30) has been separated from normal TTR(Val30) by double one‐dimensional electrophoresis using polyacrylamide gel electrophoresis followed by hybrid isoelectric focusing. The procedure appears to be appropriate for identification by the screening of patients with familial amyloidotic polyneuropathy of Portuguese type I. This is the second example to support the recently expressed hypothesis (Altlandet al., Electrophoresis1986,7, 251–259) that substitutions of electrically neutral amino acids (e. g.methionine for valine at position 30) in a polypeptide chain (e. g.the TTR monomer) may be detected by isoelectric focusing under denaturing conditions (e. g.hybrid isoelectric focusing in the presence of 8 M urea, 50 mM dithiothreitol) when the exchange is close to a charged amino acid (e. g.histidine at position 31, pI(TTR) = 5.7, pKa(His) = 6

ISSN:0173-0835    

DOI:10.1002/elps.1150071110

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

年代:1986

数据来源: WILEY

摘要:

AbstractA demountable polyacrylamide gel system is described that uses commercial components and can be adapted to any slab system. The use of GeldBond PAG film produces polyacrylamide gels that can be easily handled and allows separation of the conditions needed for polymerization from those for electrophoresis. Polymerization is performed with one common protocol, and the gel is disassembled. Gels can then be soaked or stored in any desired solutions, followed by reassembly of the equilibrated gel and electrophoresis. This procedure allows for a wide range of electrophoretic conditions starting with a single type of polymerized gel as well as permitting use of systems that have technical difficulties under normal cirumstances such as horizontal gels or those containing the cationic detergent, cetyltrimethylammonium bromide.

ISSN:0173-0835    

DOI:10.1002/elps.1150071111

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

年代:1986

数据来源: WILEY