scholarly journals Detection of anti- Keyhole limpet hemocynin (anti-KLH) in rats by double immunodiffusion (Ouchterlony) technique. v1 (protocols.io.bjs3kngn)

protocols.io ◽  
2020 ◽  
Author(s):  
Angel Justiz
Keyword(s):  
Double Immunodiffusion

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.

scholarly journals Atypical disseminated cutaneous histoplasmosis in an immunocompetent child, caused by an "aberrant" variant of Histoplasma capsulatum var. capsulatum

1999 ◽  
Vol 41 (3) ◽  
pp. 195-202 ◽  
Author(s):  
Carlos da Silva LACAZ ◽  
Gilda Maria Barbaro DEL NEGRO ◽  
Mônica Scarpelli Martinelli VIDAL ◽  
Elisabeth Maria HEINS-VACCARI ◽  
Roseli Freitas dos SANTOS ◽  
...  
Keyword(s):  
Histoplasma Capsulatum ◽  
Complement Fixation ◽  
Double Immunodiffusion ◽  
Granulomatous Reaction ◽  
Cutaneous Lesions ◽  
Biopsy Material ◽  
Specific Staining ◽  
Hyperimmune Serum ◽  
Circulating Antigen ◽  
Immunoenzymatic Assay

A case of atypical disseminated cutaneous histoplasmosis in a five-year old, otherwise healthy child, native and resident in São Paulo metropolitan area is reported. Cutaneous lesions were clinically atypical. Histologic examination disclosed a granulomatous reaction but no fungal structures could be demonstrated by specific staining nor by immunohistochemical reaction. The fungus was isolated from biopsy material on two different occasions, confirming diagnosis of an unusual fungal infection. The fungus, originally thought to be a Sepedonium sp. due to the large sized, hyaline or brownish colored tuberculated macroconidia and to lack of dimorphism (yeast form at 37 °C) produce H and M antigens, visualized by the immunodiffusion with rabbit anti-Histoplasma capsulatum hyperimmune serum. Patient’s serum sample was non reactive with H. capsulatum antigen by immunodiffusion, counterimmunoelectrophoresis and complement fixation tests, and immunoenzymatic assay failed to detect the specific circulating antigen. This serum was tested negative by double immunodiffusion when antigen obtained from one of the isolated samples was used. Both cultures were sent to Dr. Leo Kaufman, Ph.D. (Mycoses Immunodiagnostic Laboratory, CDC-Atlanta/USA), who identified them as H. capsulatum by the exoantigen and gen-probe tests. Both clinic and mycologic characteristics of the present case were atypical, suggesting the fungus isolated is an “aberrant variant” of H. capsulatum var. capsulatum, as described by SUTTON et al. in 199719. Treatment with itraconazole 100 mg/day led to cure within 90 days

Carcinoembryonic Antigen in Gynecologic Cancers. Immunohistochemical Localization and Serum Levels

1983 ◽  
Vol 69 (1) ◽  
pp. 23-30 ◽  
Author(s):  
Józef Kula ◽  
Antonina Harlozinska ◽  
Roman Richter ◽  
Zygmunt Albert ◽  
Józef Sward ◽  
...  
Keyword(s):  
Carcinoembryonic Antigen ◽  
Genital Tract ◽  
Female Genital Tract ◽  
Serum Levels ◽  
Immunohistochemical Localization ◽  
Double Immunodiffusion ◽  
Preoperative Serum ◽  
Serum Cea ◽  
Different Content ◽  
Female Genital

The presence of carcinoembryonic antigen (CEA)-dependent fluorescence was observed in almost 89% of female genital tract cancers irrespective of their histologic type. Anti-CEA serum was free of antibody to normal cross-reacting antigen. The high percentage of positive fluorescence tests did not correlate with the preoperative serum CEA levels. Double immunodiffusion tests showed different content of CEA and normal cross-reacting antigen in individual specimens of genital tract cancers.

"Microgel diffusion blotting" for sensitive detection of antibodies to extractable nuclear antigens

1990 ◽  
Vol 36 (2) ◽  
pp. 337-339 ◽  
Author(s):  
F De Keyser ◽  
G Verbruggen ◽  
E M Veys ◽  
J Nimmegeers ◽  
L Schatteman ◽  
...  
Keyword(s):  
Rheumatic Diseases ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Sensitive Detection ◽  
Nitrocellulose Membrane ◽  
Double Immunodiffusion ◽  
Simple Diffusion ◽  
Gel Layer ◽  
Nuclear Antigens

Abstract A fast immunoblotting procedure, termed "microgel diffusion blotting," is used to detect and identify antibodies to extractable nuclear antigens (i.e., to Sm, RNP, and SSB) in patients with rheumatic diseases. The method differs from the standard immunoblotting techniques by the use of ultra-thin microgels for polyacrylamide gel electrophoresis: the very thin gel layer allows transfer of proteins to a nitrocellulose membrane by simple diffusion. Principal advantages of this variant technique are its simplicity, rapidity, and reproducibility--characteristics that make the test suitable for routine application. We compared the sensitivity of the test with that of double immunodiffusion in agarose for the evaluation of humoral antinuclear immunity. Microgel diffusion blotting detected antibodies in serum at concentrations less than 0.001 of those detectable by immunodiffusion.

scholarly journals The comparison in double immunodiffusion and western blotting methods of anti-SS-A/B antibodies in patients with Sjoegren's syndrome

2003 ◽  
Vol 26 (2) ◽  
pp. 74-79
Author(s):  
Zhiquan Xu ◽  
Ken Takeuchi ◽  
Ran Matsudaira ◽  
Yoshinori Kanai ◽  
Yoshiaki Tokano ◽  
...  
Keyword(s):  
Western Blotting ◽  
Double Immunodiffusion ◽  
Sjoegren’S Syndrome

scholarly journals Serological grouping ofTreponema hyodysenteriae

1990 ◽  
Vol 105 (1) ◽  
pp. 79-85 ◽  
Author(s):  
D. J. Hampson ◽  
J. R. L. Mhoma ◽  
B. G. Combs ◽  
J. I. Lee
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Vaccine Development ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cross Reactivity ◽  
Low Molecular Weight ◽  
Serological Response ◽  
Double Immunodiffusion ◽  
Sds Page ◽  
Treponema Hyodysenteriae

SUMMARYTwo Australian isolates ofTreponema hyodysenteriaewhich did not fit within the current serological grouping system for these bacteria wrere examined by agarose gel double immunodiffusion tests (AGDP). Isolate Vic1 was serologically unique, and we propose that it becomes the type organism for a new sixth serological group ofT. hyodysenteriae(Group F). Isolate Q1 was unusual in that lipopolysaccharide (LPS) extracted from it reacted strongly in AGDP with serum raised against the type organism for serogroup D (A1), and also weakly with serum raised against the type organism for serogroup B (WA1). The nature of this cross-reactivity was examined by using cross-absorbed antisera in AGDP, and by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis.The pattern of serological cross-reactivity between Q1, A1 and WA1 was complex and was not fully defined, but the isolate Q1 apparently shared low molecular weight ‘serogroup’ LPS antigens with A1, and shared higher molecular weight LPS antigens with WA1. On this basis Q1 was designated as belonging to serogroup D, although it was recommended that this be qualified as D (B) to indicate the presence of weak cross-reactivity with serogroup B. Such serological cross-reactivity may have significance in relation to the development of immunity toT. hyodysenteriae. Isolate Q1 may be a potentially useful organism for vaccine development because of its ability to induce a good serological response to LPS of treponemes from both serogroups D and B.

IDENTIFICATION AND PURIFICATION OF A NEW PROTEIN FROM WATER-SOLUBLE EXTRACTS OF HUMAN ADRENAL GLAND

1979 ◽  
Vol 82 (3) ◽  
pp. 383-NP ◽  
Author(s):  
M. A. AL-AWQATI ◽  
Y. B. GORDON ◽  
T. CHARD
Keyword(s):  
Molecular Weight ◽  
Adrenal Gland ◽  
Gel Filtration ◽  
Water Soluble ◽  
Double Immunodiffusion ◽  
Molecular Weights ◽  
Physicochemical Methods ◽  
Two Peaks ◽  
Sepharose 4B ◽  
New Protein

An homogenate of human foetal adrenal gland was subjected to negative immunoabsorption by column chromatography using anti-whole human serum coupled to Sepharose 4B. Two peaks were eluted and used to immunize rabbits. The antisera produced were absorbed and tested for specificity by double immunodiffusion. Two antigens, which appeared to be specific to the adrenal gland, were identified having molecular weights of 25 000 and 65 000 as determined by gel filtration. The lower molecular weight antigen was isolated by physicochemical methods and found to be a protein. The amino acid composition is reported.

scholarly journals Purification and some properties of cytosolic cobalamin-binding protein in Euglena gracilis

1987 ◽  
Vol 247 (3) ◽  
pp. 679-685 ◽  
Author(s):  
F Watanabe ◽  
Y Nakano ◽  
S Kitaoka
Keyword(s):  
Euglena Gracilis ◽  
Binding Proteins ◽  
Microsomal Fraction ◽  
Binding Protein ◽  
Physiological Role ◽  
Binding Activity ◽  
Thiol Groups ◽  
Double Immunodiffusion ◽  
Ph Dependency ◽  
Ks Values

In Euglena gracilis SM-ZK, a bleached mutant of E. gracilis z, the cobalamin- (Cbl-)binding activity was distributed in cytosol (49.2%), mitochondria (20.3%) and microsomal fraction (20.4%). The cytosolic Cbl-binding activity gave two major peaks in isoelectric focusing. The Cbl-binding protein with pI 3.2 was purified 6500-fold in a yield of 19.9%, and that with pI 4.7 5800-fold in a yield of 11.9%. The monomeric Mr values of both Cbl-binding proteins were about 66,000. The Cbl-binding activity of both proteins showed a very low pH-dependency, and thiol groups and metal ions were not concerned with the Cbl-binding activities. The Ks values of the Cbl-binding proteins with pI 3.8 and 4.7 for CN-Cbl were 1.0 and 2.0 nM respectively. The Cbl-binding protein with pI 3.8 was shown to be immunologically identical with the protein with pI 4.7 by double-immunodiffusion experiments against antibody to the protein with pI 3.8. The two cytosolic Cbl-binding proteins did not show the activities of Cbl-dependent enzymes in E. gracilis, N5-methyltetrahydrofolate:homocysteine methyltransferase, methylmalonyl-CoA mutase and ribonucleotide reductase, suggesting that the two cytosolic Cbl-binding proteins play a physiological role as intracellular Cbl carriers.

scholarly journals Structure of O-Polysaccharide and Lipid A of Pantoea Agglomerans 8488

Biomolecules ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 804 ◽  
Author(s):  
Tetiana V. Bulyhina ◽  
Evelina L. Zdorovenko ◽  
Ludmila D. Varbanets ◽  
Alexander S. Shashkov ◽  
Alexandra A. Kadykova ◽  
...  
Keyword(s):  
Lipid A ◽  
Fatty Acid Analysis ◽  
Complex Compounds ◽  
Pantoea Agglomerans ◽  
Ionization Mass ◽  
Two Dimensional ◽  
Double Immunodiffusion ◽  
Toxic Activity ◽  
Serological Activity ◽  
Mild Acid

The Pantoea agglomerans 8488 lipopolysaccharide (LPS) was isolated, purified and characterized by monosaccharide and fatty acid analysis. The O-polysaccharide and lipid A components of the LPS were separated by mild acid degradation. Lipid A was studied by electrospray ionization mass spectrometry (ESI-MS) and found to consist of hexa-, penta-, tetra- and tri-acylated species. Two-dimensional nuclear magnetic resonance (NMR) spectroscopy revealed the following structure of the O-polysaccharide repeating unit →3)-α-L-Rhap-(1→6)-α-D-Manp-(1→3)-α-L-Fucp-(1→3)-β-D-GlcNAcp-(1→. The LPS showed a low level of toxicity, was not pyrogenic, and reduced the adhesiveness index of microorganisms to 2.12, which was twofold less than the control. LPS modified by complex compounds of germanium (IV) and tin (IV) were obtained. It was found that six LPS samples modified by Sn compounds and two LPS samples modified by Ge compounds lost their toxic activity when administered to mice in a dose of LD50 (105 µg/mice or 5 mg/kg). However, none of the modified LPS samples changed their serological activity in an Ouchterlony double immunodiffusion test in agar.

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