Identification of the German Cockroach Allergens in Korean Atopy Using SDS - PAGE and Western Blot Analysis

In the present study, lung, trachea and serum samples from broiler flocks slaughtered at an abattoir in Elazig province located in the East of Turkey were examined for the presence of Ornithobacterium rhinotracheale using culture and enzyme-linked immunosorbent assay (ELISA). The identity was latter proved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot analysis, and polymerase chain reaction (PCR) assays. A total of 324 serum and 250 lung and trachea samples were collected from 10 commercially reared chicken flocks showing respiratory manifestations. The samples were obtained from different flocks. The causative agent (ORT) was isolated from trachea (1.5%) of five chickens and from both lung and trachea (0.4%) of only one chicken in the bacteriological examination of tissues. The presence of antibodies against ORT was detected in 33 (10.2%) of the 324 sera by ELISA. A 784 bp fragment of the 16S rRNA gene was amplified using specific primers in the PCR. All ORT isolates that were positive by culture were also detected to be positive by the PCR. SDS-PAGE protein profiles of whole cell extracts showed a high similarity for all the isolates with a major band of the molecular weight of 33 kDa (kiloDalton). Results of Western blot analysis indicate four antigenic fractions predominantly with molecular weights of 33, 42, 52 and 66 kDa.

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Identification of important allergens in German cockroach extracts by sodium dodecylsulfate–polyacrylamide gel electrophoresis and Western blot analysis

Journal of Allergy and Clinical Immunology ◽

10.1016/s0091-6749(95)70132-x ◽

1995 ◽

Vol 95 (4) ◽

pp. 877-885 ◽

Cited By ~ 24

Author(s):

Jonathan J. Musmand ◽

W.Elliott Horner ◽

Manuel Lopez ◽

Samuel B. Lehrer

Keyword(s):

Western Blot ◽

Blot Analysis ◽

Gel Electrophoresis ◽

Western Blot Analysis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecylsulfate ◽

German Cockroach ◽

Polyacrylamide Gel

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SDS-PAGE and Western blot analysis of triactinomyxon spores of Myxobolus cerebralis, the cause of whirling disease in salmonid fish

Journal of Fish Diseases ◽

10.1046/j.1365-2761.2003.00497.x ◽

2003 ◽

Vol 26 (10) ◽

pp. 621-625 ◽

Cited By ~ 1

Author(s):

H Soliman ◽

K Geissler ◽

M El-Matbouli

Keyword(s):

Western Blot ◽

Blot Analysis ◽

Western Blot Analysis ◽

Salmonid Fish ◽

Whirling Disease ◽

Myxobolus Cerebralis ◽

Sds Page

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Syringe Purification and HRP-Conjugation of Goat IgGs Used in Quality Control of Erythropoietin

Journal of Applied Biotechnology ◽

10.5296/jab.v6i2.12829 ◽

2018 ◽

Vol 6 (2) ◽

pp. 11

Author(s):

Marcel Bassil ◽

Nataliia Pavliuchenko ◽

Elia Raya

Keyword(s):

Quality Control ◽

Horseradish Peroxidase ◽

Western Blot ◽

Affinity Chromatography ◽

Blot Analysis ◽

Western Blot Analysis ◽

Protein G ◽

Sds Page

Goat anti-rabbit IgGs were purified from serum using Protein G affinity chromatography. The purity of eluted fractions was tested using SDS-PAGE, and the functionality of IgGs was tested in Western Blot analysis of Erythropoietin, after conjugation with Horseradish Peroxidase.

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A meiotic DNA polymerase from a mushroom, Agaricus bisporus

Biochemical Journal ◽

10.1042/bj2990335 ◽

1994 ◽

Vol 299 (2) ◽

pp. 335-340 ◽

Cited By ~ 13

Author(s):

K Takami ◽

S Matsuda ◽

A Sono ◽

K Sakaguchi

Keyword(s):

Western Blot ◽

Dna Polymerase ◽

Blot Analysis ◽

Western Blot Analysis ◽

Agaricus Bisporus ◽

Polymerase Activity ◽

Polymerase Beta ◽

Optimal Activity ◽

Sds Page ◽

Molecular Masses

A meiotic DNA polymerase [DNA nucleotidyltransferase (DNA-directed), EC 2.7.7.7], which likely has a role in meiotic DNA repair, was isolated from a mushroom, Agaricus bisporus. The purified fraction displays three bands in SDS/PAGE, at molecular masses of 72 kDa, 65 kDa and 36 kDa. Optimal activity is at pH 7.0-8.0 in the presence of 5 mM Mg2+ and 50 mM KCl and at 28-30 degrees C, which is the temperature for meiosis. This enzyme is resistant to N-ethylmaleimide and sensitive to 2′,3′-dideoxythymidine 5′-triphosphate, suggesting that it is a beta-like DNA polymerase. These characteristics are similar to those of Coprinus DNA polymerase beta [Sakaguchi and Lu (1982) Mol. Cell. Biol. 2, 752-757]. In Western-blot analysis, the antiserum against the Coprinus polymerase reacts only with the 65 kDa band, which coincides with the molecular mass of the Coprinus polymerase. Western-blot analysis also showed that the antiserum could react with crude extracts not only from the Agaricales family, to which Agaricus and Coprinus belong, but also from different mushroom families and Saccharomyces. The Agaricus polymerase activity can be found only in the meiotic-cell-rich fraction, but the enzyme is also present in the somatic cells in an inactive state.

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Interactions of Streptococcus mutansFimbria-Associated Surface Proteins with Salivary Components

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.6.3.400-404.1999 ◽

1999 ◽

Vol 6 (3) ◽

pp. 400-404 ◽

Cited By ~ 8

Author(s):

Chad A. Ray ◽

Linda E. Gfell ◽

Tiffany L. Buller ◽

Richard L. Gregory

Keyword(s):

Western Blot ◽

Blot Analysis ◽

Western Blot Analysis ◽

Salivary Protein ◽

Enzyme Linked Immunosorbent Assay ◽

Major Protein ◽

Sds Page ◽

Whole Saliva ◽

Coated Surfaces ◽

Fimbrial Protein

ABSTRACT Streptococcus mutans has been implicated as the major causative agent of human dental caries. S. mutans binds to saliva-coated tooth surfaces, and previous studies suggested that fimbriae may play a role in the initial bacterial adherence to salivary components. The objectives of this study were to establish the ability of an S. mutans fimbria preparation to bind to saliva-coated surfaces and determine the specific salivary components that facilitate binding with fimbriae. Enzyme-linked immunosorbent assay (ELISA) established that the S. mutans fimbria preparation bound to components of whole saliva. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot techniques were used to separate components of whole saliva and determine fimbria binding. SDS-PAGE separated 15 major protein bands from saliva samples, and Western blot analysis indicated significant binding of the S. mutans fimbria preparation to a 52-kDa salivary protein. The major fimbria-binding salivary protein was isolated by preparative electrophoresis. The ability of the S. mutans fimbria preparation to bind to the purified salivary protein was confirmed by Western blot analysis and ELISA. Incubation of the purified salivary protein with the S. mutans fimbria preparation significantly neutralized binding of the salivary protein-fimbria complex to saliva-coated surfaces. The salivary protein, whole saliva, and commercial amylase reacted similarly with antiamylase antibody in immunoblots. A purified 65-kDa fimbrial protein was demonstrated to bind to both saliva and amylase. These data indicated that the S. mutans fimbria preparation and a purified fimbrial protein bound to whole-saliva-coated surfaces and that amylase is the major salivary component involved in the binding.

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The identification of a reaction site of glutathione mixed-disulphide formation on gammaS-crystallin in human lens

Biochemical Journal ◽

10.1042/bj20031367 ◽

2004 ◽

Vol 379 (3) ◽

pp. 595-600 ◽

Cited By ~ 28

Author(s):

Jane CRAGHILL ◽

Andrew D. CRONSHAW ◽

John J. HARDING

Keyword(s):

Western Blot ◽

Blot Analysis ◽

Western Blot Analysis ◽

Outer Part ◽

Human Lens ◽

Reaction Site ◽

Lens Proteins ◽

Reducing Conditions ◽

Sds Page

The glutathionylation of human lens proteins was examined by Western-blot analysis with an anti-GSH antibody and scanning. Several different glutathionylated proteins were observed, and a 47 kDa band was of particular interest. This band did not appear after SDS/PAGE under reducing conditions, suggesting that it was a glutathionylated fraction. The 47 kDa band was found principally in the outer part of the lens, the cortex, but not in the lens nucleus where older proteins are present. The 47 kDa component was composed of βB1-, βB2- and γS-crystallin, with the γS-crystallin having glutathione bound at Cys-82 and at Cys-22, Cys-24 or Cys-26. We conclude that when glutathione becomes bound to γS-crystallin, it causes it to bind in turn to the β-crystallin polypeptides to form a dimer.

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Transformation Studies of Bacillus thuringiensis cryIC Gene into a Nitrogen-Fixing Azospirillum lipoferum

Zeitschrift für Naturforschung C ◽

10.1515/znc-2001-3-412 ◽

2001 ◽

Vol 56 (3-4) ◽

pp. 245-248 ◽

Cited By ~ 1

Author(s):

Ramalingam Gounder ◽

Narayanan Rajendran

Keyword(s):

Bacillus Thuringiensis ◽

Carbon Source ◽

Western Blot ◽

Blot Analysis ◽

Western Blot Analysis ◽

Carbon Sources ◽

Toxin Gene ◽

Nitrogen Fixing ◽

Azospirillum Lipoferum ◽

Sds Page

AbstractA lepidopteran toxin gene, cryIC (pSB607) from entom opathogenic Bacillus thuringiensis subsp. aizawai was introduced into nitrogen-fixing A zospirillum lipoferum by transformation. Regeneration of spheroplasts was achieved at 99% with 39% frequency of regeneration. Transformants were screened on NB kanamycin with ampicillin plates and 4 transformants were selected after ten generations. SDS-PAGE and Western blot analysis confirmed the presence of a 68 kDa protein in the transformants. Studies on utilization of carbon sources indicate that glucose and sucrose are the most favorable carbon sources and 2 % molasses is the cheap alternate carbon source for the better growth of parent A. lipoferum and transformants.

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