摘要:

AbstractIn this series of papers, we describe the computer system we have developed for two‐dimensional (2‐D) gel analysis. Here the acquisition part of the system is described and a set of criteria able to measure the overall performance of the system is presented. These criteria are essentially based on frequency domain considerations using Fourier transforms. We show that the transfer function characteristics of the input system must be carefully checked to avoid any image distortion; we propose an easily computed criterion to obtain setting conditions giving a correct system response. These features are discussed considering the specificity of the 2‐D gel images that can be well modeled by a multi‐Gaussian surface. Then the visualization device and its main abilities to treat and modify the aspect of the images are presented. Statistical functions using second‐order image histograms are introduced. They can be used to look at the general aspect of the images, as a global and accurate criterion for quality checking. For this purpose, covariance, entropy and inertia have been found to be most suitable. Second‐order histograms give additional information on image anisotropy and can direct the user towards the convenient values of the parameters for the subsequent treatments. Coding is considered from both the point of view of image compression and image sending throughout a computer network. An efficient code protecting all the image informations is suggested an

ISSN:0173-0835    

DOI:10.1002/elps.1150070802

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

年代:1986

数据来源: WILEY

摘要:

AbstractThis work describes the techniques used to detect the presence of a spot at a given place in a two‐dimensional gel and to compute its volume. This is done using filtering techniques to extract significant information from rough images. Four types of treatments were applied to images in order to improve the detection of spots belonging to different categories. Mathematical techniques of morphology were used to lower the effect of spot overlapping, while histogram modifications using power functions were used to improve the detection of major and minor spots, respectively, thereby generating four processed images. On each generated image, spot detection is computed by looking at the local maxima. Then the four spots lists are gathered and the volume is computed according to a spot model. To improve the accuracy of volume detection, a spot model constituted by two half‐Gaussian curves in each space direction is used. The pitch of each spot according to the horizontal direction is also computed. A total of 8 parameters, including spot position, shape parameters, surface intensity and volume, is gathered into a spot list which constitutes the reduced information which will be handled subsequently. The choice of a model allows us to get a round the improper volume calculations caused by spot overlapping. The mutual penetration of one spot into another is thus automatically taken into account. A set of statistical considerations is computed from these data in order to validate our spot model, to check for parameter independence, to obtain quality criteria which may be used to judge gel quality, to propose different diagnoses for the observed defaults and to lead to a self‐adaption process of treatment parameters. These statistical considerations can be used to study the effect of each type of treatment and to compare the effects of each filter on the final r

ISSN:0173-0835    

DOI:10.1002/elps.1150070803

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

年代:1986

数据来源: WILEY

摘要:

AbstractMononuclear leukocytes of the human peripheral blood were separated by counter‐current centrifugal elutriation. In this way a group of lymphocytes was isolated, which contained B‐lymphocytes with an electrophoretic mobility (EM) of 0.9 × 10−4(cm2V−1s−1) and T‐lymphocytes with an EM of 1.1 × 10−4(cm2V−1s−1). This group of lymphocytes was incubated for 7 days in the presence of monocytes and pokeweed mitogen. Then the culture supernatants were screened. Non‐specific human antibodies were found in all culture supernatants. In 20% of the supernatants human antibodies against sheep red blood cells could be detected. The viable cells of the cultures were separated into 5 fractions by free flow electrophoresis. Then the cells of the different fractions were tested for their capability to produce antibodies. The tests revealed that antibody‐secreting cells had an EM around 0.9 × 10−4(cm2V−1s−1) and that the greater part of the cells, isolated from pokeweed mitogen cultures, had a significant higher EM. Thus free flow electrophoresis could be applied to enrich ant

ISSN:0173-0835    

DOI:10.1002/elps.1150070804

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

年代:1986

数据来源: WILEY

摘要:

AbstractFree flow electrophoresis was used for the purification of human tissue plasminogen activator (t‐PA) produced by recombinant yeast cells. This method was employed for three principal reasons: (a) yeast t‐PA purified by immunoaffinity chromatography was not pure enough to carry out protein chemical characterization, (b) the amount of yeast t‐PA available for purification was extremely limited, (c) t‐PA has a strong tendency to adsorb to support material; a support‐free separation method therefore seemed to be a reasonable way to further purify the enzyme. Free flow electrophoresis was first performed using solutions containing the soluble proteins of yeast cell extract, and secondly, using t‐PA fractions prepared by immunoaffinity chromatography, Yeast t‐PA purified by immunoaffinity chromatography and then by free flow electrophoresis is more than 80% pure and suitable for characterization experiments such as glycosylation studies or N‐terminal sequenc

ISSN:0173-0835    

DOI:10.1002/elps.1150070805

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

年代:1986

数据来源: WILEY

摘要:

AbstractA high resolution agarose gel system (Panagel) with silver staining and polyacrylamide disc gel electrophoresis were used to demonstrate oligoclonal bands in cerebrospinal fluid (CSF) from 150 patients identified as having either inflammatory neurological diseases (110 patients) or non‐inflammatory neurological diseases (40 patients). The detection of oligoclonal band patterns in high resolution agarose electrophoresis is sensitive: 65% (polyacrylamide gel electrophoresis: 50%) and specific: 95% (polyacrylamide gel electrophoresis: 73%) with respect to inflammatory neurological diseases. This procedure is easier to perform than polyacrylamide disc gel electrophoresis and is recommended for routine examination of CSF especially for the detection of oligoclonal band

ISSN:0173-0835    

DOI:10.1002/elps.1150070806

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

年代:1986

数据来源: WILEY

摘要:

AbstractImmunoblotting after sodium dodecyl sulfate polyacrylamide gel electrophoresis of the purified third component of human complement (C3) and its fragments and of unpurified C3 and its degradation fragments in serum or plasma has been examined. The pattern of C3 degradation observed corresponded well with previous observations made with purified proteins. The immunoblotting technique demonstrated a high resolution, and proved to be capable of providing a full analysis of mixtures of C3 fragments in serum with the use of chain‐fragment‐specific antibody preparations. Additional heterogeneity in C3 and its breakdown products were observed, and covalent adducts between activated C3 and other proteins were detected directly in serum and plasma without any purification of the components. The results further indicated that antibodies raised against denatured C3 reacted quantitatively and qualitatively better with the nitrocellulose‐bound C3 than those antibodies raised to native C3, probably reflecting the paratially denatured state of molecules after separation by polyacrylamide gel electrophoresis and blotting onto the nitrocell

ISSN:0173-0835    

DOI:10.1002/elps.1150070807

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

年代:1986

数据来源: WILEY

摘要:

AbstractThe method presented was developed specifically to be used for the electrophoretic analysis of polymeric proteins in order to establish the number of aggregates and that of their subunits. Electrophoresis is carried out on a single slab; without sodium dodecyl sulfate (SDS) in the first dimension and with SDS in the second dimension. The method is possible since for one‐dimensional SDS gel electrophoresis, SDS is only required in the cathode vesse

ISSN:0173-0835    

DOI:10.1002/elps.1150070808

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

年代:1986

数据来源: WILEY

摘要:

AbstractA novel method is described to serve as a positive control by including an antiserum raised against and reacted with blots of the molecular standards used in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The marker antiserum may find further application on nylon based adsorptive membranes which cannot be stained by conventional method

ISSN:0173-0835    

DOI:10.1002/elps.1150070809

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

年代:1986

数据来源: WILEY

摘要:

AbstractA new technique is presented for the detection of human galactokinase from crude hemolysate by the method of polyacrylamide gel isoelectric focusing. The method utilizes low sample volume and can be completed within 5 h. One gel can be used to screen 15 samples, making the procedure ideal for screening erythrocyte galactokinase in newborn screening programs. This method can be ideal for many laboratories interested in screening for galactokinase variants.

ISSN:0173-0835    

DOI:10.1002/elps.1150070810

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

年代:1986

数据来源: WILEY

摘要:

AbstractRectangular sections of leaves, vacuum‐infiltrated with distilled water or aqueous solutions of potassium chloride, were applied directly onto the surface of polyacrylamide gels followed by isoelectric focusing. Soluble peroxidases (free and ionically bound) were released from the tissue on isoelectric focusing with resultant characteristic zymograms for the different plant tissue

ISSN:0173-0835    

DOI:10.1002/elps.1150070811

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

年代:1986

数据来源: WILEY