Microzone electrophoresis of the unconcentrated and concentrated CSF

1978 ◽  
Vol 218 (1) ◽  
pp. 73-76 ◽  
Author(s):  
H. Glasner
Keyword(s):  
Microzone Electrophoresis

Screening for Albuminuria: A Case for Estimation of Albumin in Urine

1975 ◽  
Vol 21 (12) ◽  
pp. 1795-1798 ◽  
Author(s):  
E BenGershôm
Keyword(s):  
Total Protein ◽  
Urinary Albumin ◽  
Green Method ◽  
Green Is ◽  
Protein Estimation ◽  
Microzone Electrophoresis ◽  
Bromcresol Green

Abstract The usefulness of bromcresol green for estimating albumin in urine was evaluated by comparison with the Laurell "rocket" technique. In contrast to the bromcresol green method applied for urinary albumin, rather doubtful results were obtained with conventional (Microzone) electrophoresis for albumin and with precipitation techniques for total protein estimation. Albumin estimation with bromcresol green is recommended as a more reliable substitute for total-protein estimations in urine. Limitations of bromcresol green are also pointed out.

Effects of Temperature on Serum Protein Components in the Killifish, Fundulus heteroclitus

1970 ◽  
Vol 27 (2) ◽  
pp. 404-409 ◽  
Author(s):  
Bruce L. Umminger
Keyword(s):  
Immune Response ◽  
Cellulose Acetate ◽  
Serum Protein ◽  
Fundulus Heteroclitus ◽  
Serum Proteins ◽  
High Mobility ◽  
Electrophoretic Patterns ◽  
Effects Of Temperature ◽  
Protein Components ◽  
Microzone Electrophoresis

Electrophoretic patterns of serum proteins were studied in Fundulus heteroclitus acclimated to temperatures from 30 to −1.5 C. Microzone electrophoresis on cellulose acetate revealed eight distinct zones (seven anodal and one cathodal to the origin) at all temperatures. These zones were grouped into three larger zones (I, II, and III) for convenience in analysis. Low mobility proteins of zone III decreased in the cold to a greater degree than did high mobility proteins of zones II and I. The greater reduction in these low mobility proteins, which are comparable to human gamma globulins, might produce a less active immune response in cold-acclimated killifish.

scholarly journals RELATIONSHIPS BETWEEN RELATIVE BINDING AFFINITY AND ELECTROPHORETIC BEHAVIOR OF RABBIT ANTIBODIES TO STREPTOCOCCAL CARBOHYDRATES

1971 ◽  
Vol 133 (3) ◽  
pp. 424-441 ◽  
Author(s):  
K. Eichmann ◽  
J. Greenblatt
Keyword(s):  
Binding Affinity ◽  
Variable Region ◽  
Reciprocal Relationship ◽  
Binding Affinities ◽  
Igg Antibody ◽  
Intravenous Injections ◽  
Relative Binding Affinity ◽  
Relative Binding ◽  
Electrophoretic Behavior ◽  
Microzone Electrophoresis

After repeated intravenous injections with Group C streptococcal vaccine, most rabbit antisera were shown to contain one or more IgG antibody components, as revealed by microzone electrophoresis. A procedure for the fractionation of multiple IgG antibody components from such streptococcal antisera is described. Separation is achieved on the basis of differences in relative binding affinities of the antibody components to immunoabsorbent columns. The evidence suggests that the electrophoretic mobility, and thus the net charge of an antibody, bears a reciprocal relationship to its binding affinity for the streptococcal Group C antigens. Furthermore, the relative binding affinity affords another means to assess the functional homogeneity of streptococcal antibodies. A possible relationship between light chain variable-region subclasses and binding affinities of streptococcal antibodies is discussed.

scholarly journals ENZYME ACTIVITIES IN CHICK EMBRYONIC CARTILAGE

1971 ◽  
Vol 49 (2) ◽  
pp. 459-467 ◽  
Author(s):  
C. Arsenis ◽  
R. Eisenstein ◽  
L. W. Soble ◽  
K. E. Kuettner
Keyword(s):  
Enzyme Activities ◽  
High Speed ◽  
Subcellular Fraction ◽  
Subcellular Fractionation ◽  
Enzymatic Activities ◽  
Sensitive Technique ◽  
Embryonic Cartilage ◽  
Microzone Electrophoresis ◽  
High Speed Supernatant ◽  
Supernatant Solution

Some characteristic enzymatic activities were determined in chick embryonic cartilage and compared with the analogous activities in bone and liver. Chondrocytes were isolated, broken by sonication, and subjected to subcellular fractionation to yield a nuclear pellet, the mitochondrial, lysosomal, and microsomal fractions, and the high speed supernatant solution. It was established that these fractions are characterized by enzymatic activities usually associated with similar fractions in other tissues, but with some quantitative differences. Lysozyme, a particulate-associated enzyme in other tissues, was not detected in any subcellular fraction even by the sensitive technique of microzone electrophoresis and is therefore considered to be primarily extracellular in cartilage.

Evaluation of the Microzone System for Simultaneous Electrophoresis of 16 Sera

1971 ◽  
Vol 17 (1) ◽  
pp. 34-36 ◽  
Author(s):  
Benjamin Fingerhut ◽  
Antonia Ortiz
Keyword(s):  
Cellulose Acetate ◽  
Cellulose Acetate Membrane ◽  
Large Numbers ◽  
Sample Application ◽  
Electrophoresis System ◽  
Microzone Electrophoresis ◽  
Abnormal Results

Abstract We investigated the feasibility of using the "Microzone" electrophoresis system to produce 16 instead of eight electrophorograms on a single cellulose—acetate membrane by placing eight specimens at the left of center, rotating the cellulose—acetate membrane and supporting bridge 180° horizontally, and again applying eight samples to the left of center. The membrane, now with eight samples each on anodic and cathodic sides, is electrophoresed according to the usual recommended procedure. There was a statistically demonstrated association between sample application position and results (for 96 samples). This technique is nevertheless useful for determining albumin/globulin ratios and for screening large numbers of samples. Sera with borderline or abnormal results should be rerun by the usual technique.

Purification and Properties of the Thrombin-Like Principle of Agkistrodon acutus Venom and Its Comparison with Bovine Thrombin

1971 ◽  
Vol 26 (02) ◽  
pp. 224-234 ◽  
Author(s):  
Chaoho Ouyang ◽  
Jau-Shyong Hong ◽  
Che-Ming Teng
Keyword(s):  
Specific Activity ◽  
Ph Value ◽  
Activate Factor ◽  
Clot Retraction ◽  
Crude Venom ◽  
Sephadex Column ◽  
Bovine Thrombin ◽  
Heat Stable ◽  
Purification And Properties ◽  
Microzone Electrophoresis

SummaryBy means of DEAE-Sephadex column chromatography, Agkistrodon acutus venom was separated into 12 fractions. The thrombin-like activity was concentrated in Fr.10. This fraction was rechromatographed on Sephadex G-200. A single band was found on microzone electrophoresis at pH 7.4. A single boundary with S20, w of 3.82 S, which was almost symmetrical, was observed by ultracentrifugation. The estimated molecular weight is 33,500. The chemical analysis shows that the thrombin-like principle of the venom is a glycoprotein. The specific activity is 13 times higher than that of the crude venom. The optimal pH value of the thrombin-like principle (pH 7.5) of the venom is almost identical with that of the bovine thrombin (pH 7.2). The thrombinlike principle of the venom is not affected by heparin, while the clotting activity of the bovine thrombin is inhibited by heparin. The thrombin-like principle of the venom is much more heat stable than the bovine thrombin.No clot retraction was found with the thrombin-like principle of the venom, while the marked clot retraction occurred after plasma was coagulated with thrombin.Thrombin can activate factor XIII (Fibrin stabilizing factor), while the thrombinlike principle can not.

scholarly journals A SERUM FACTOR IN CHRONIC HYPOCOMPLEMENTEMIC NEPHRITIS DISTINCT FROM IMMUNOGLOBULINS AND ACTIVATING THE ALTERNATE PATHWAY OF COMPLEMENT

1974 ◽  
Vol 139 (5) ◽  
pp. 1249-1261 ◽  
Author(s):  
Enrique H. Vallota ◽  
Otto Götze ◽  
Hans L. Spiegelberg ◽  
Judith Forristal ◽  
Clark D. West ◽  
...  
Keyword(s):  
Sedimentation Coefficient ◽  
Sucrose Density Gradient ◽  
Normal Serum ◽  
Chronic Glomerulonephritis ◽  
Complement Pathway ◽  
Classical Complement Pathway ◽  
Alternate Pathway ◽  
Alternate Complement Pathway ◽  
Microzone Electrophoresis ◽  
C5a Anaphylatoxin

Nephritic factor (C3NeF) has been isolated from plasma of patients with hypocomplementemic chronic glomerulonephritis (HCG) by ion exchange and molecular sieve chromatography. This material was further treated with solidified anti-Ig antiserum. The purified material failed to react with antiserum to human IgG, IgG3, Fab, Fc, and kappa and lambda chains, but retained full C3NeF activity. The nonidentity of C3NeF with IgG was further demonstrated by Ouchterlony analysis using anti-IgG and anti-C3NeF. Isolated C3NeF was found to be a protein with a sedimentation coefficient of 7S and a mol wt of 150,000 daltons, which on microzone electrophoresis and gel electrophoresis at pH 8.6 behaved as a γ-globulin. C3NeF is not a C1q precipitin and does not activate the classical complement pathway. Unlike cobra venom factor, it failed to enter into a complex with C3 proactivator (C3PA) when incubated with normal human serum (NHS) and then subjected to sucrose density gradient ultracentrifugation. The action of isolated C3NeF on C3 requires C3PA, C3PA convertase (C3PAse), and properdin (P). Similarly, C3PA conversion by C3NeF requires P, C3PAse, and C3. Total hemolytic activity was lost by incubation of 64 µg of C3NeF/1 ml NHS at 37°C for 30 min. Both C3a and C5a anaphylatoxin could be generated by C3NeF in serum previously depleted of anaphylatoxin inactivator. Anti-C3NeF was found to detect an antigen in all NHS tested. Treatment of NHS with solidified anti-C3NeF caused impairment of the alternate complement pathway. It failed to sustain lysis of glutathione-treated human erythrocytes initiated by inulin. It is conceivable that the normal serum constituent which is removed by anti-C3NeF constitutes the inactive precursor of C3NeF, and a heretofore unrecognized component of the alternate pathway.

scholarly journals ISOLATION AND IMMUNOCHEMICAL CHARACTERIZATION OF RABBIT 7S ANTI-IGG WITH RESTRICTED HETEROGENEITY

1973 ◽  
Vol 137 (6) ◽  
pp. 1354-1368 ◽  
Author(s):  
Viktor A. Bokisch ◽  
Jen Wei Chiao ◽  
David Bernstein
Keyword(s):  
Binding Site ◽  
Light Chain ◽  
Guinea Pigs ◽  
Potent Inhibitor ◽  
Molecular Heterogeneity ◽  
Antibody Specificity ◽  
Immunochemical Characterization ◽  
Microzone Electrophoresis ◽  
Antigen Antibody

7S anti-IgGs were isolated from four rabbit antistreptococcal antisera by the use of immunoabsorbent columns. The isolated proteins were of restricted molecular heterogeneity; they formed a monodisperse band on microzone electrophoresis and had a limited number of light chain bands when analyzed on urea polyacrylamide gel. The binding site of the 7S anti-IgGs was detected in the F(ab')2 portion of the molecule. The binding site has antibody specificity for the Fc portion of IgG. For one 7S anti-IgG the combining site on the Fc portion could further be defined. A pepsin fragment of Fc, described as Pep-III', was a potent inhibitor of the coprecipitation of 7S anti-IgG with antigen-antibody complexes. An idiotypic cross-reaction was detected between the 7S and 19S anti-IgGs isolated from the same rabbit with anti-idiotype sera prepared in guinea pigs. This idiotypic specificity was not detected in the 7S anti-IgGs of 20 other rabbits.

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