ISSN:0173-0835    

DOI:10.1002/elps.1150030502

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

年代:1982

数据来源: WILEY

摘要:

AbstractIn order to increase accuracy and precision of quantifying one‐dimensional electropherograms by digital signal processing, a set of algorithms is presented that operate on the optical density values acquired by a linear scanning densitometer. These operations include smoothing of the measuring values, background correction by an optimized piecewise linear baseline, and integration of the peak areas after two‐sided fit of half‐Gaussian profiles to each stained protein band. The algorithms are implemented as BASIC‐programs and are readily adaptable to a variety of electrophoretic separations. The procedure has been applied successfully in research [1] as well as in clinical evaluati

ISSN:0173-0835    

DOI:10.1002/elps.1150030503

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

年代:1982

数据来源: WILEY

摘要:

AbstractRecently we reported about a linear correlation between the logarithm of the size of native proteins (log mol mass or log Stokes' radius) and the square root of their migration distance (‐ √D) in linear polyacrylamide (PAA)‐gradient gels (G. M. Rothe and H. Purkhanbaba, Electrophoresis 1982,3, 33–42). The linearity between log MM and √D is not subject to time using homogeneous buffers in electrophoresis, no matter how the constants of the corresponding regression lines, slope and intercept change as a function of time. The realiability of this correlction has been re‐examined with 0.7 mm thin gel plates and extending the time of electrophoresis under non‐denaturating conditions from 2 to 95 h. It is demonstrated that the linear log size ‐ √D‐relationship is also applicable to sodium dodecyl sulfate (SDS) linear PAA gradient

ISSN:0173-0835    

DOI:10.1002/elps.1150030504

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

年代:1982

数据来源: WILEY

摘要:

AbstractAn isotachophoretic procedure for the estimation of the electrophoretic mobility of ions is described. The parameter, called apparent mobility (AM), is a function of the conditions of assay. Defining a relative apparent mobility (RAM) as the percentage value of the AM of the ion studied and the AM of a fully dissociated reference substance, an operational estimate of the electrophoretic mobility of ionic species is obtained. RAM values, therefore, are only a function of the leading ion and of the ion studied. If this ion demonstrates acid‐base equilibria, its RAM value will become pH dependent. As RAM values are independent on the conditions of assay, classification is meaningful. For a number of anions, RAM values may help to find out the optimal conditions for separation of ions by analytical isotachophoresi

ISSN:0173-0835    

DOI:10.1002/elps.1150030505

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

年代:1982

数据来源: WILEY

摘要:

AbstractA method is reported for the separation and partial purification of proteins on a preparative scale (around 100 mg or more total protein per separation run), using the isotachophoretic principle. The technique was carried out using a granulated gel (Ultrodex) as supporting medium, with conventional flat‐bed electrophoresis equipment. This method was applied to the separation of multiple forms of glutathione S‐transferase, derived from the soluble cytosolic fraction of cat liver homogenate. Of particular interest in the present work was the isolation of the highest mobility form of the enzyme, which also functions as a cytosolic ligand‐binding protein for bilirubin. The advantages and disadvantages of isotachophoresis are compred with those of isoelectric focusing, using a simliar apparative set‐up. The latter had the primary drawback that some forms of the enzyme, particularly those with most acidic or alkaline isoelectric points, were, to a certain extent, irreversibly inactivated following focusing, whereas isotachophoresis yielded considerably higher volume activities and resulted in less inactivation during separation. The loss in activity of some forms of the enzyme was particularly marked in isoelectric focusing in thinner gels with agarose as supporting phase. Isotachophoresis has become associated with quite complicated instrumental requirements. The simple form of this technique reported in the present work should provide a useful addition to the spectrum of methods available for protein separation and purification. The major limitation of the flat‐bed configuration is that the actual resolution obtained cannot be fully exploited owing to the technical difficulties associated with sectioning the

ISSN:0173-0835    

DOI:10.1002/elps.1150030506

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

年代:1982

数据来源: WILEY

摘要:

AbstractRecycling isoelectric focusing (RIEF) was used to isolate monoclonal antibodies from an electrophoretically heterogeneous rabbit antibody population. Milligram quantities of antibodies were processed to single band purity in 4 h. The ease of operation, extremely short run times, and high resolution of the RIEF apparatus give this system great flexibility. The demonstrated capacity of the RIEF system to process large quantities of protein suggests that the system is applicable to the largescale purification of antibodies.

ISSN:0173-0835    

DOI:10.1002/elps.1150030507

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

年代:1982

数据来源: WILEY

摘要:

AbstractIsoelectric focusing was used to study the antibody clonotype patterns of outbred New Zealand White rabbits during pronlonged immunization with the gram positive bacteriumMicrococcus lysodeikticus.Antisera were examined by analytical isoelectric focusing in 1% agarose gels using inexpensive carrier ampholytes which were synthesized in the laboratory. Clonotype patterns of antimicrococcal antibodies were visualized by radioautography using [125I]‐labeled micrococcal carbohydrate antigen. Examination of the antibody clonotype patterns in antisera from a number of outbred rabbits revealed that variations in clonotype expression often occurred within an individual during successive rounds of immunization. These studies demonstrate the utility of analytical isoelectric focusing for monitoring antibody clonotype changes within an individual during immunizatio

ISSN:0173-0835    

DOI:10.1002/elps.1150030508

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

年代:1982

数据来源: WILEY

摘要:

AbstractProteins in the cytosol, post‐nuclear particulate and nuclear fractions from dorsolateral and ventral prostates of rats were analyzed and compared by sodium dodecyl sulfate (SDS)‐polyacrylamide gel electrophoresis. Protein species separated in the gel were stained with Coomassie Brilliant Blue R‐250 (CBB), and expressed as (apparent molecular weight of the respective species x 10−3)K. (i) In the cytosol fraction the contents of 30K, 67K, 110K, and 120K proteins were abundant in the dorsolateral prostate. On the other hand, 14K and 16K proteins were specific to the ventral prostate. By carbohydrate staining, major glycoproteins were found to be 120K and 16K for the dorsolateral and ventral prostates, respectively. (ii) In post‐nuclear particulate fractions in both lobes, proteins having molecular weights greater than 42 000 were abundant. Of these, the contens of 65K, 105K and 120K were higher in the dorsolateral prostate than in the ventral prostate. Glycoproteins having the same apparent molecular weight as detected in the cytosol fractions were also found in the post‐nuclear particulate fractions of both lobes. (iii) In the nuclear fraction H2A, H2B, H3 and H4 histones were fundamentally similar between the two lobes, whereas H1 histones including two subspecies were slightly but significantly higher in the ventral prostate than the dorsolateral prostate. The most remarkable difference between the lobes was the content of 20K in the dorsolateral prostate which was at least 20 times higher than that found in the ventr

ISSN:0173-0835    

DOI:10.1002/elps.1150030509

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

年代:1982

数据来源: WILEY

摘要:

AbstractImmuno‐labeling of human peripheral blood lymphocytes with either polyglutaraldehyde or polyacrolein microspheres was shown by analytical electrophoresis to generate two electrokinetically distinct sub‐populations of lymphocytes. By means of free‐flow electrophoresis, the high mobility fraction was found to consist predominantly of T lymphocytes using an anti‐T cell antibody, while the low mobility fraction consisted of lymphocytes labeled with immunomicrospheres, identifying these cells as B lymphocytes. The immunomicrosphere‐induced reduction of B‐cell mobility below that of the T cells thus allowed the enrichment and recovery of viable T cells in preparative quantities by free‐flow el

ISSN:0173-0835    

DOI:10.1002/elps.1150030510

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

年代:1982

数据来源: WILEY

ISSN:0173-0835    

DOI:10.1002/elps.1150030511

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

年代:1982

数据来源: WILEY