摘要:

AbstractThe relation between the electrophoretic migration velocity and the concentration of the non‐ionic detergent Triton X‐100 in the buffer was studied by crossed immunoelectrophoresis of hydrophilic and amphiphilic proteins. The migration velocity of hydrophilic human plasma proteins was not affected by the presence of Triton X‐100 in the agarose gels in the concentration range of 1–0 % v/v. In contrast, amphiphilic proteins, like the human erythrocyte proteins glycophorin, band 3 protein and acetylcholinesterase, and the plasma high density lipoprotein (HDL) apolipoprotein, showed increasing migration velocity with decreasing detergent concentration in the gels. An increase of 20–80 % was observed for different amphiphilic proteins when the Triton X‐100 concentration was lowered from 1 % to 0.03 % v/v. This probably reflects the influence of the size of the bound detergent micelle on the charge density of the protein. Thus, by performing corssed immunoelectrophoresis with different concentrations of Triton X‐100 in the first‐dimensional gel it is possible to identify amphiphilic proteins. Presence of 1 % v/v Triton X‐100 in the second‐dimensional gel ensures reliable identification

ISSN:0173-0835    

DOI:10.1002/elps.1150070502

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

年代:1986

数据来源: WILEY

摘要:

AbstractAffinity electrophoresis was applied to determine the dissociation constants and the thermodynamic parameters for the interactions between the mouse myeloma, MOPC 315, and trinitrophenyl (Tnp) and dinitrophenyl (Dnp) haptens. The myeloma protein consists of monomeric, dimeric, and trimeric forms of IgA. The dimer had a higher affinity to the haptens than the monomer, but its Fab' fragment had lower affinity than the monomer. All of these myeloma proteins had higher affinities to Tnp‐hapten than to Dnp‐hapten. The affinity of MOPC 315 proteins increased when the temperature increased. The van't Hoff plots for all of these interactions gave straight lines within the temperature range from 7 °C to 40 °C. The ΔH° were calculated by using the linear van't Hoff plots. The ΔS° were calculated from the D̊H° and D̊G°. The D̊H° and D̊S° were both positive. Hence, the interactions were endothermi

ISSN:0173-0835    

DOI:10.1002/elps.1150070503

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

年代:1986

数据来源: WILEY

摘要:

AbstractA method is described which allows the preparation of retinae, laminae, opticallobes, protocerebrum and proboscis of single heads ofDrosophila. The protein content of these tissues was examined with sodium dodecyl sulfate‐polyacrylamide gel electrophoresis followed by Coomassie Brilliant Blue R‐250 and glycoprotein staining. A catalog of 39 major proteins or protein subunits was obtained. Their relative proportions were determined quantitatively for the different tissues fromDrosophilawildtype strains AS and C‐S as well as the mutants

ISSN:0173-0835    

DOI:10.1002/elps.1150070504

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

年代:1986

数据来源: WILEY

摘要:

AbstractA method for separating fetal hemoglobin Sardinia (AγT, a variant containing threonine in position 75 instead of isoleucine) from normal fetal hemoglobin is described, based on the use of shallow immobilized pH gradients (pH 7.30–7.55). The novelty of the fractionation technique is the direct analysis of intact, native hemoglobin tetramers, while present methods utilize only reverse‐phase, high performance liquid chromatography of denatured, heme‐free globin

ISSN:0173-0835    

DOI:10.1002/elps.1150070505

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

年代:1986

数据来源: WILEY

摘要:

AbstractUncrosslinked polyacrylamide, a viscous liquid, provides a molecular sieving effect in electrophoresis qualitatively the same as that of crosslinked gels. This finding confirms previous results by others using uncrosslinked polyacrylamide mixed with various gel matrices. It provides a sieving medium constituted by an entirely and necessarily random association of inert fibers, as stipulated for interpretation of migration by the Ogston theory. Ferguson plots of several proteins in uncrosslinked polyacrylamide exhibitYovalues smaller and less divergent from one another than in 5 % crosslinked gels, suggesting a curvature of the Ferguson plots of crosslinked polyacrylamide gels at low %T. If linearity of these plots at low %T, 0 %C could be demonstrated, a means would be found to determine accurate values of free mobility by their linear extrapolation to 0 gel concentration. Potentially, uncrosslinked polyacrylamide at low concentrations may provide a tool for the analytical sieving of subcellular particles. However, electrophoresis in uncrosslinked polyacrylamide at concentrations of less than 10 % is perturbed by convection, and thus these potential applications will depend on combining it with a suitable anticonvective device.

ISSN:0173-0835    

DOI:10.1002/elps.1150070506

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

年代:1986

数据来源: WILEY

摘要:

AbstractA compact device, capable of being clipped onto the end of a 3 mm thick analytical polyacrylamide slab gel, has been developed for the collection of resolved protein zones as they elute from the end of the gel. Its unique feature is that eluted proteins were collected between two polyacrylamide‐embedded paper membranes and removed either by continuous or intermittent buffer elution. Because of this direct coupling to large gels identical to those used for analytical electrophoresis, protein zones were obtained with high resolution. The method was applicable to non‐denaturing cathodic and anodic electrophoresis, as well as sodium dodecyl sulfate electrophoresis. Medium‐scale quantities (up to 10 mg/cm2cross‐sectional area) of protein were fractionated in as little as 4 h with high rec

ISSN:0173-0835    

DOI:10.1002/elps.1150070507

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

年代:1986

数据来源: WILEY

摘要:

AbstractWhen screening natural populations for genetic variation in mitochondrial (mt) DNA it is necessary to screen as many individuals as possible for as large a number of base pairs as possible. This report describes a combination of laboratory techniques – isolation of mtDNA by either density ultracentrifugation or a phenol extraction procedure and DNA fragment visualization by a sensitive silver staining technique. These routines remove the major restriction of tissue sample size and number of individuals that can be screened and allows the investigation of at least 1000 base pairs from 1 g of tissue or 3–400 base pairs from 100 mg of frozen tis

ISSN:0173-0835    

DOI:10.1002/elps.1150070508

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

年代:1986

数据来源: WILEY

摘要:

AbstractRecently we described a procedure for pouring immobilized pH gradients at ambient temperature in the presence of a neutral buffer (0.1MTris‐phosphate pH 6.8) (Altland and Rossmann,Electrophoresis1985,6, 314–325). After washing in plain water and in 1 % glycerolin water, the gels were dried, sealed in polyethylene bags and stored at –20°C until use. The reproducibility of a batch of 20 gels with a linear pH 6–10 gradient, poured under computer control, was studied by rehydrating the dried gels in a solution containing 8Murea, 50 mMdithiothreitol and 0.5 % carrier ampholytes (pH 6–10) and by running human red cell lysates as samples. A comparison of the globin patterns obtained at various time intervals within a range of 7 months revealed that there was no detectable shift in the pH gradient within this period and that gels stored for 7 months can be used to clearly identify all human globins from newborns and adults. Since the tested pH gradient was prepared from Immobilines claimed as being the most sensitive to degradation by hydrolysis (e. g.Immobiline pKa8.5 and 9.3), it is concluded that storage of dry gels, rehydrated prior to use, is a realistic alternative to the preparation of fresh gels from stored stock solutions for achieving reproducible results in long‐term compara

ISSN:0173-0835    

DOI:10.1002/elps.1150070509

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

年代:1986

数据来源: WILEY

摘要:

AbstractRenouncing protein fixation with aldehydes, the polyacrylamide gels are incubated in alcoholic solutions of silver nitrate containing acetic acid at 60–70 °C. for a short time and developed with NaOH, also at 60–70 °C. Using various methods of protein fixation, the detection limits of human albumin were determined for 0.75 and 1.5 mm thick polyacrylamide gels with appropriate thickness correction. An overlay technique with silver nitrate containing polyacrylamide gels is described. Dispersing Coomassie Brilliant Blue G‐250 in a solution containing HNO3and NH4NO3allows rapid silver staining of prestain

ISSN:0173-0835    

DOI:10.1002/elps.1150070510

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

年代:1986

数据来源: WILEY

摘要:

AbstractThe sensitivity for protein detection after isoelectric focusing can be enhanced using a newly developed applicator strip with a concentration effect. This effect is achieved with funnel‐shaped slots, which are placed parallel to the pH gradient. Furthermore, sample volumes varying from 1–90 μL can be applied without distortion to the pH gradient. A comparison with other commercially available applicator strips was undertaken using the phosphoglucomutase sy

ISSN:0173-0835    

DOI:10.1002/elps.1150070511

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

年代:1986

数据来源: WILEY