Fractionation and antigenic characterization of organelles ofEimeria tenellasporozoites

Parasitology ◽  
1992 ◽  
Vol 104 (1) ◽  
pp. 1-9 ◽  
Author(s):  
U. Kawazoe ◽  
F. M. Tomley ◽  
J. A. Frazier
Keyword(s):  
Molecular Weight ◽  
Western Blotting ◽  
Sucrose Gradient ◽  
Eimeria Tenella ◽  
2Nd Generation ◽  
Antigenic Characterization ◽  
Electron Microscopical ◽  
Sucrose Gradient Ultracentrifugation ◽  
Antigenic Reactivity

SUMMARYSporozoites ofEimeria tenellawere disrupted by sonication and subcellular fractions were separated by sucrose gradient ultracentrifugation. Fractions from gradients were characterized by electron microscopical appearance and their polypeptide and antigenic profiles determined by PAGE and Western blotting with antisera to sporozoites and 1st- and 2nd-generation merozoites. Fractions containing micronemes, rhoptries or membranes showed markedly different polypeptide content and antigenic reactivity. Microneme epitopes were strongly conserved between sporozoites and 2nd-generation merozoites whereas the majority of rhoptry epitopes and many membrane epitopes were sporozoite specific. The only polypeptide of sporozoites which was strongly recognized by antisera raised to 1st generation merozoites was a microneme antigen of molecular weight approximately 100 kDa.

scholarly journals CHARACTERIZATION OF VirB4 PROTEIN OF LOCAL ISOLATE Brucella abortus WITH WESTERN BLOTTING TECHNIQUE

2018 ◽  
Vol 12 (1) ◽  
Author(s):  
Ratih Novita Praja ◽  
Didik Handijatno ◽  
Setiawan Koesdarto ◽  
Aditya Yudhana
Keyword(s):  
Molecular Weight ◽  
Western Blotting ◽  
Virulence Factor ◽  
Brucella Abortus ◽  
Protein Band ◽  
Molecular Weights ◽  
Local Isolate ◽  
Blotting Technique

This research aimed to characterize VirB4 protein of local isolate Brucella abortus with Western blotting method. The result showed that there were four protein bands with molecular weights of 64.61, 59.25, 21.63, and 16.70 kDa by triggering a reaction between the whole Brucella abortus and anti-Brucella abortus serum. The results also revealed that there was only one protein band with a molecular weight of 59.25 kDa triggering a reaction between the whole Brucella abortus and anti-VirB Brucella abortus serum. Finally, it can be concluded that VirB4 protein can affect the virulence factor of Brucella abortus, successfully characterized with the appearance of one band with a molecular weight of 59.25 kDa by using Western blotting method.

CONGENITAL GOITRE IN SHEEP: ISOLATION OF THE IODOPROTEINS WHICH REPLACE THYROGLOBULIN

1976 ◽  
Vol 71 (2) ◽  
pp. 179-192 ◽  
Author(s):  
C. E. DOLLING ◽  
B. F. GOOD
Keyword(s):  
Molecular Weight ◽  
Thyroid Gland ◽  
Sucrose Gradient ◽  
Sedimentation Coefficient ◽  
Agarose Gel ◽  
Merino Sheep ◽  
Thyroid Glands ◽  
Sucrose Gradient Ultracentrifugation ◽  
Australian Merino

SUMMARY Immunological and chromatographic studies of proteins from the congenital goitre of South Australian Merino sheep revealed that normal thyroglobulin is absent from the thyroid glands of these sheep. However, a thyroglobulin-immunoreactive iodoprotein was isolated by affinity chromatography on agarose gel to which antibody against thyroglobulin had been covalently bound. Sucrose-gradient ultracentrifugation indicated that this iodoprotein had a sedimentation coefficient of 8S and a molecular weight of approximately 175000. This iodoprotein is therefore about one quarter the size of normal thyroglobulin (19S; 660000) and is similar in size to the subunit of thyroglobulin (3-8S; 165000) although this has usually been described as non-iodinated except when derived by reductive fission. In addition the goitre extract contained iodoproteins which had the immunological properties of serum albumin and immunoglobulin G. Determination of the iodine and iodoamino acid content of the hydrolysed iodoproteins revealed that they contained iodothyronines which were able to contribute to the production of thyroid hormones although the total iodothyronine content of the goitrous gland was less than that of the normal sheep thyroid gland.

Partial characterization of a non-proteinaceous, low molecular weight antigen of Eimeria tenella

2000 ◽  
Vol 86 (6) ◽  
pp. 461-466
Author(s):  
John R. Barta ◽  
Shan A. Tennyson ◽  
Marco L. Schito ◽  
Harry D. Danforth ◽  
Donald S. Martin
Keyword(s):  
Molecular Weight ◽  
Eimeria Tenella ◽  
Partial Characterization ◽  
Low Molecular Weight

Etching of Polymers by Oxygen Plasma

1968 ◽  
Vol 26 ◽  
pp. 408-409
Author(s):  
Virgil Peck ◽  
W. L. Carter
Keyword(s):  
Molecular Weight ◽  
Oxygen Plasma ◽  
Flow Time ◽  
Microscopical Study ◽  
Surface Layers ◽  
Organic Vapors ◽  
Sample Position ◽  
Accuracy Of Measurement ◽  
Bulk Polymers ◽  
Electron Microscopical

Any electron microscopical study of the morphology of bulk polymers has throughout the years been hampered by the lack of any real ability to produce meaningful surface variations for replication. True etching of polymers should show crystalline and amorphous regions in some form of relief. The use of solvents, acids, organic vapors, and inert ion bombardment to etch samples has proved to be useful only in limited applications. Certainly many interpretations of these results are subject to question.The recent use of a radiofrequency (R. F.) plasma of oxygen to degrade and remove organic material with only minor heating has opened a new possibility for etching polymers. However, rigid control of oxygen flow, time, current, and sample position are necessary in order to obtain reproducible results. The action is confined to surface layers; the molecular weight of the polymer residue after heavy etching is the same as the molecular weight of the polymer before attack, within the accuracy of measurement.

Isolation and Characterization of Plasminogen Activator from Pig Leucocytes

1974 ◽  
Vol 31 (01) ◽  
pp. 072-085 ◽  
Author(s):  
M Kopitar ◽  
M Stegnar ◽  
B Accetto ◽  
D Lebez
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Ph Optimum ◽  
Isolation And Characterization ◽  
Ph Range ◽  
Inhibition Studies ◽  
Final Purification

SummaryPlasminogen activator was isolated from disrupted pig leucocytes by the aid of DEAE chromatography, gel filtration on Sephadex G-100 and final purification on CM cellulose, or by preparative gel electrophoresis.Isolated plasminogen activator corresponds No. 3 band of the starting sample of leucocyte cells (that is composed from 10 gel electrophoretic bands).pH optimum was found to be in pH range 8.0–8.5 and the highest pH stability is between pH range 5.0–8.0.Inhibition studies of isolated plasminogen activator were performed with EACA, AMCHA, PAMBA and Trasylol, using Anson and Astrup method. By Astrup method 100% inhibition was found with EACA and Trasylol and 30% with AMCHA. PAMBA gave 60% inhibition already at concentration 10–3 M/ml. Molecular weight of plasminogen activator was determined by gel filtration on Sephadex G-100. The value obtained from 4 different samples was found to be 28000–30500.

Studies on the Characterization of Factor VIII and a Co-factor VIII

1974 ◽  
Vol 31 (02) ◽  
pp. 328-338
Author(s):  
M. M. P Paulssen ◽  
H. L. M. A Vandenbussche-Scheffers ◽  
P. B Spaan ◽  
T de Jong ◽  
M. C Planje
Keyword(s):  
Molecular Weight ◽  
Factor Viii ◽  
The Body ◽  
Haemophilia A ◽  
Von Willebrand's Disease ◽  
Von Willebrand’S Disease ◽  
Polysaccharide Content ◽  
Plasma Factor ◽  
Two Factors

SummaryFactor VIII occurs in the body in two different forms. In lymph factor VIII is bound to chylomicra. In plasma, factor VIII is bound to a protein.After delipidation of chylomicra we obtained a glycoprotein with a high polysaccharide content and a molecular weight of approx. 160,000.In plasma, factor VIII is attached to a protein which is present in normal concentrations in plasma of patients with haemophilia A and in serum (co-factor VIII).This factor is deficient in both the plasma and the serum of patients with von Willebrand’s disease.The binding between factor VIII and co-factor VIII is reversible.Some properties of these two factors are described.

Cross-Linked αsγt-Chain Hybrids in Plasma Clots Studied by 1D- and 2D Electrophoresis and Western Blotting

1993 ◽  
Vol 70 (03) ◽  
pp. 438-442 ◽  
Author(s):  
B Grøn ◽  
C Filion-Myklebust ◽  
S Bjørnsen ◽  
P Haidaris ◽  
F Brosstad
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Human Plasma ◽  
Western Blotting ◽  
Isoelectric Point ◽  
Factor Xiii ◽  
Cross Linking ◽  
2D Electrophoresis ◽  
Complex Phenomenon ◽  
Fibrinopeptide A

SummaryFibrinogen and fibrin related chains in reduced human plasma as well as the bonds interlinking partially cross-linked fibrin from plasma clots have been studied by means of 1D- and 2D electrophoresis and Western blotting. Immunovisualization of reduced plasma or partially cross-linked fibrin with monoclonal antibodies specific for the α-chains or the γ-chains have shown that several bands represent material belonging to both chains. In order to decide whether these bands constitute αγ-chain hybrids or superimposed α- and γ-chain dimers, the cross-linked material was separated according to both isoelectric point (pI) and molecular weight (MW) using Pharmacia’s Multiphor II system. Western blotting of the second dimension gels revealed that partially cross-linked fibrin contains αsγt-chain hybrids and γ- polymers, in addition to the well-known γ-dimers and α-polymers. The main αsγt-chain hybrid has a pI between that of the α- and the γ-chains, a MW of about 200 kDa and contains Aα-chains with intact fibrinopeptide A (FPA). It was also observed that soluble fibrinogen/fibrin complexes as well as partially cross-linked fibrin contain degraded α-dimers with MWs close to the γ-dimers. These findings demonstrate that factor XIII-catalyzed cross-linking of fibrin is a more complex phenomenon than earlier recognized.

Rapid Isolation of High Molecular Weight Urokinase from Native Human Urine

1982 ◽  
Vol 47 (03) ◽  
pp. 197-202 ◽  
Author(s):  
Kurt Huber ◽  
Johannes Kirchheimer ◽  
Bernd R Binder
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Human Urine ◽  
Gel Filtration ◽  
Chain Structure ◽  
The Other ◽  
Single Chain ◽  
Apparent Homogeneity ◽  
Sequential Adsorption

SummaryUrokinase (UK) could be purified to apparent homogeneity starting from crude urine by sequential adsorption and elution of the enzyme to gelatine-Sepharose and agmatine-Sepharose followed by gel filtration on Sephadex G-150. The purified product exhibited characteristics of the high molecular weight urokinase (HMW-UK) but did contain two distinct entities, one of which exhibited a two chain structure as reported for the HMW-UK while the other one exhibited an apparent single chain structure. The purification described is rapid and simple and results in an enzyme with probably no major alterations. Yields are high enough to obtain purified enzymes for characterization of UK from individual donors.

Prothrombin Metz : Purification and Characterization of a Variant of Human Prothrombin

1979 ◽  
Author(s):  
M Ribieto ◽  
J Elion ◽  
D Labie ◽  
F Josso
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Factor Xa ◽  
Polyacrylamide Gel ◽  
Cleavage Pattern ◽  
Purification And Characterization ◽  
Double Immunodiffusion ◽  
The Family

For the purification of the abnormal prothrombin (Pt Metz), advantage has been taken of the existence in the family of three siblings who, being double heterozygotes for Pt Metz and a hypoprothrombinemia, have no normal Pt. Purification procedures included barium citrate adsorption and chromatography on DEAE Sephadex as for normal Pt. As opposed to some other variants (Pt Barcelona and Madrid), Pt Metz elutes as a single symetrical peak. By SDS polyacrylamide gel electrophoresis, this material is homogeneous and appears to have the same molecular weight as normal Pt. Comigration of normal and abnormal Pt in the absence of SDS, shows a double band suggesting an abnormal charge for the variant. Pt Metz exhibits an identity reaction with the control by double immunodiffusion. Upon activation by factor Xa, Pt Metz can generate amydolytic activity on Bz-Phe-Val-Arg-pNa (S2160), but only a very low clotting activity. Clear abnormalities are observed in the cleavage pattern of Pt Metz when monitored by SDS gel electrophoresis. The main feature are the accumulation of prethrombin l (Pl) and the appearance of abnormal intermediates migrating faster than Pl.

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