scholarly journals SDS-PAGE Characterisation of Crude Seed and Leaf Proteins in Corchorus Species

2015 ◽  
Vol 7 (2) ◽  
pp. 177-183
Author(s):  
Christiana Adeyinka AJALA ◽  
Joseph Akintade MORAKINYO
Keyword(s):  
Protein Separation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Electrophoretic Study ◽  
Single Linkage ◽  
Banding Patterns ◽  
Leaf Proteins ◽  
Sds Page ◽  
Single Linkage Cluster ◽  
Linkage Cluster

Crude protein separation was carried out for Corchorus incisifolious, Corchorus aestuans, Corchorus tridens and Corchorus olitorious using Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE). Plants were collected both from wild and cultivated sites and samples included leaves and seeds for the electrophoretic study. Distinct polymorphism in electrophoretic banding patterns of seed and leaf proteins following SDS- PAGE was observed in the four Corchorus species studied. Forty- two polypeptide bands were observed in the seed and a total of eleven polypeptide bands were observed in the leaves of the Corchorus species studied. The electrophoretic study revealed protein bands with various intensities ranging from high, to low and faint. The results showed that there was variation in both the seed and leaf proteins of the Corchorus species studied. A dendrogram constructed based on the Single Linkage Cluster Analysis (SLCA) clustering method revealed three major clusters for seeds. Cluster I consisted of C. incisifolious and C. aestuans, cluster II consisted of C. tridens, while cluster III consisted of C. olitorious. The leaf protein extracts were grouped into two clusters, cluster one containing C. incisifolious and C. aestuans, while the other contained C. tridens and C. olitorious.

Application of metaproteomic analysis for studying extracellular polymeric substances (EPS) in activated sludge flocs and their fate in sludge digestion

2008 ◽  
Vol 57 (12) ◽  
pp. 2009-2015 ◽  
Author(s):  
C. Park ◽  
R. F. Helm
Keyword(s):  
Activated Sludge ◽  
Protein Separation ◽  
Extracellular Polymeric Substances ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cation Exchange Resin ◽  
Methanosarcina Barkeri ◽  
Extracellular Proteins ◽  
Sludge Digestion ◽  
Sds Page

Metaproteomic analysis, comprising protein separation and identification, was applied to study extracellular proteins in activated sludges and to track their fate in sludge digestion under both anaerobic and aerobic conditions. The complex sludge proteins were first separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and further analysed by liquid chromatography tandem mass spectrometry (LC-MS/MS) to search their identification. Base extraction and cation exchange resin (CER) method were used to extract EPS from sludges at 0, 12 and 30 days of batch digestion. Several important observations were made during this study. Firstly, protein bands were well separated by extraction/SDS-PAGE protocol used in this study. Secondly, numerous protein bands remained after digestion, indicating that these proteins are not easily degradable in sludge digestion. Thirdly, protein bands detected following anaerobic and aerobic digestion differed, suggesting that proteins degraded in two different digestion environments are not the same. Finally, protein bands that emerged distinctively following anaerobic digestion was found to be subunits of methyl-coenzyme M reductase, the enzyme involved in methane generation, in Methanosarcina barkeri. These results demonstrated that metaproteomic investigation on activated sludge EPS is useful for studying floc formation in activated sludges and their degradation in various digestion environments.

scholarly journals Migration Behavior of Immunoglobulin Y in Native Page Electrophoresis

2019 ◽  
Vol 2 (1) ◽  
pp. 50-56
Author(s):  
Umul Karimah
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Protein Separation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sodium Deoxycholate ◽  
Migration Behavior ◽  
Immunoglobulin Y ◽  
Native Page ◽  
Nonreducing Condition ◽  
Sds Page

ABSTRACT Protein separation with electrophoresis in native state (native Polyacrylamide Gel Electrophoresis, native PAGE) is rarely applied in laboratory which results in limited scientific report. Native PAGE result can be used in further assay analysis or shows important protein-protein interaction. This study aimed to investigate immunoglobulin Y(IgY) behavior in separation using native PAGE. IgY protein is used since many properties of IgY are already known. IgY was isolated from follicles yolk and examined using clear native PAGE (CN-PAGE) and Sodium Dodecyl Sulphate (SDS) PAGE, both in reducing and nonreducing condition. CN-PAGE electropherogram of IgY showed streaked band from ~409-1048 kDa indicating IgY aggregation. Whereas SDS-PAGE in reducing condition showed normal result, ~68 kDa and ~18,7 kDa protein bands correlated with heavy and light chain of IgY respectively. Moreover SDS PAGE in nonreducing condition also resulted in normal IgY band sized ~185 kDa. Dilution and disaggregation using sodium dodecyl sulphate (SDS), sodium deoxycholate (SDC), sarcosyl, urea, and glycerol did not affect the migration behavior. We concluded that IgY is not in aggregate form. Migration behavior of IgY in CN-PAGE which was slow, produced streaked band, and appeared to have very high molecular weight is possibly due to low electronegativity (pI = 6,6±0,9) in CN-PAGE condition (buffer pH 8,3) and planar conformation of IgY. Keywords: electrophoresis, immunoglobulin Y, native PAGE.

Study on Mechanism of Cross-Linking of Peanut Protein Isolate Modified with Transglutaminase

2012 ◽  
Vol 550-553 ◽  
pp. 1304-1308 ◽  
Author(s):  
Qing Jie Sun ◽  
Liu Xiong ◽  
Xiang Hui Bu ◽  
Yan Liu
Keyword(s):  
Ir Spectra ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Isolate ◽  
Peanut Protein ◽  
Cross Linking ◽  
Banding Patterns ◽  
Peanut Protein Isolate ◽  
Sds Page ◽  
Ft Ir

The mechanism of cross-linking of peanut protein isolate (PPI) modified with transglutaminase was investigated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier Transformation Infrared (FT-IR) spectra. SDS-PAGE banding patterns indicated that the contents of arachin and conarachin after transglutaminase (TGase) modification were decreased and high molecular weight polymers were formed. SDS-PAGE banding patterns also suggested that cross-linking effects were accomplished in the presence of transglutaminase and the main component participating in cross-linking was arachin. The representative FT-IR spectra of arachin, conarachin modified with TGase treatment appeared the sharp peak at 1680~1630cm-1 region, which showed that intramolecular cross-linking was occurred, respectively. Compared with FT-IR spectra of arachin, conarachin modified with TGase treatment, the spectra of PPI modified with TGase treatment appeared two characteristic absorption at 1546.29cm-1 and 1330.75cm-1, suggesting that cross-linking was occurred between arachin and conarachin and the ε-(γ-glutamyl) isopeptide bond generated.

scholarly journals Identification of barley cultivars using SDS-PAGE electrophoresis

1992 ◽  
Vol 1 (1) ◽  
pp. 73-82
Author(s):  
Janne Roininen ◽  
Eero Nissilä ◽  
Matti Puolimatka ◽  
Seppo Pulli
Keyword(s):  
Seed Storage Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Seed Storage ◽  
Hordeum Vulgare L ◽  
Banding Patterns ◽  
Breeding Lines ◽  
Barley Cultivars ◽  
Sds Page ◽  
Single Seeds

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was applied to cultivar identification. Three different extractions methods were used to extract and fractionate the seed storage protein subunits from crushed single seeds of barley (Hordeum vulgare L.). Fifty-four genotypes including breeding lines and released cultivars were analysed and grouped according to the variation found in their protein banding patterns. In the first extraction, eight genotypes showed unique hordein subunit composition whilst remainder fell into 11 groups of 2 to 8. The other two extractions were carried out to characterize those genotypes producing identical banding patterns when using the first method. Relative mobility (REM) values for hordein bands were determined. Genetic background was found to strongly effect the determination of hordein composition of barley genotypes. Those genotypes with largely common ancestry showed often similar hordein composition and were difficult to identify whereas genotypes possessing unique hordein banding patterns had clearly exceptional pedigree. The effect of the row-type on hordein banding pattern was not clear as both two-row and many-row barleys were found to produce identical patterns. Intra-cultivar hordein polymorfism was found in three cultivars.

scholarly journals Leaf Protein Electrophoresis and Taxonomy of Species of Jatropha L. (Euphorbiaceae)

2012 ◽  
Vol 4 (3) ◽  
pp. 92-96
Author(s):  
Olaniran Temitope OLADIPO ◽  
Herbert Chukwuma ILLOH
Keyword(s):  
Protein Profile ◽  
Protein Electrophoresis ◽  
Leaf Protein ◽  
Relative Mobility ◽  
Single Linkage ◽  
Banding Patterns ◽  
Taxonomic Importance ◽  
Young Leaves ◽  
Single Linkage Cluster ◽  
Linkage Cluster

The systematic relationship existing among members of the all important genus Jatropha was studied using leaf protein electrophoresis. The aim was to identify possible taxonomic importance of the protein profile in the estimation and elucidation of the taxonomic affinity of the six species of Jatropha (Jatropha curcas Linn., J. podagrica Hook., J. gossypifolia Linn., J. mutifida Linn., J. tanjorensis Ellis & Saroja and J. integerrima Linn.) found in Nigeria. The species were screened for total protein banding patterns using gel electrophoresis. Young leaves (0.8 g) of the plants were washed with distilled water and macerated with sterile mortar and pestle in 0.8% Phosphate Buffer-Saline (PBS) containing 0.4 M NaCl at pH 8.0. Results reveal that protein banding pattern was taxon specific. Generic band occurs at 8.3. The highest number of interspecific bands (4) exists between J. podagrica and J. multifida. Variations exist not only in the number of bands but also in the intensity of the bands. Sokal and Sneath coefficient of similarity ranges between 11.1-44.4 %. Single linkage Cluster Analysis (SLCA) of the relative mobility values of the protein in the taxa shows partial agreement with current sub generic and sectional delimitation of the species based on morphology and anatomy of the species.

Electrophoretic Heterogeneity of Lipopolysaccharides of Actinobacillus actinomycetemcomitans

1988 ◽  
Vol 67 (3) ◽  
pp. 574-576 ◽  
Author(s):  
C.I. Hoover
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Epidemiological Studies ◽  
Actinobacillus Actinomycetemcomitans ◽  
Relative Mobility ◽  
Banding Patterns ◽  
Sds Page ◽  
Staining Techniques ◽  
Serotype B ◽  
Phenol Water

Electrophoretic banding patterns of lipopolysaccharides (LPS), as observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), have proved to be useful in studies relating LPS structure to virulence and as epidemiological markers. In this report, LPS of Actinobacillus actinomycetemcomitans from outer membrane fractions and hot phenol-water extracts were analyzed by SDS-PAGE and LPS-specific silver-staining techniques. Both intra- and inter-strain heterogeneity of A. actinomycetemcomitans LPS was observed. Twelve strains of A. actinomycetemcomitans, representative of the three described serotypes, were assigned to LPS subtypes based on the relative mobility of their most rapidly migrating LPS band. All three serotype a strains (29523, aB75, and GA3), two (29524 and SAC11A) of five serotype b strains, and two (aB67 and SAC5A) of four serotype c strains were assigned to LPS subtype I. The three remaining serotype b strains (29522, Y4, and JP2) were assigned to LPS subtype II, and the remaining two serotype c strains (SAC6A and SAC12A) were assigned to LPS subtype III. LPS subtyping may serve as an adjunct or alternative to serotyping in epidemiological studies.

Heterozygous Abnormal Fibrinogen Osaka III with the Replacement of γ Arginine-275 by Histidine Has an Apparently Higher Molecular Weight γ-Chain Variant

1992 ◽  
Vol 68 (05) ◽  
pp. 534-538 ◽  
Author(s):  
Nobuhiko Yoshida ◽  
Shingi Imaoka ◽  
Hajime Hirata ◽  
Michio Matsuda ◽  
Shinji Asakura
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Tetraacetic Acid ◽  
Fibrin Monomer ◽  
Sds Page ◽  
Thrombotic Tendency ◽  
Chain Variant

SummaryCongenitally abnormal fibrinogen Osaka III with the replacement of γ Arg-275 by His was found in a 38-year-old female with no bleeding or thrombotic tendency. Release of fibrinopeptide(s) by thrombin or reptilase was normal, but her thrombin or reptilase time in the absence of calcium was markedly prolonged and the polymerization of preformed fibrin monomer which was prepared by the treatment of fibrinogen with thrombin or reptilase was also markedly defective. Propositus' fibrinogen had normal crosslinking abilities of α- and γ-chains. Analysis of fibrinogen chains on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the system of Laemmli only revealed the presence of abnormal γ-chain with an apparently higher molecular weight, the presence of which was more clearly detected with SDS-PAGE of fibrin monomer obtained by thrombin treatment. Purified fragment D1 of fibrinogen Osaka III also seemed to contain an apparently higher molecular weight fragment D1 γ remnant on Laemmli gels, which was digested faster than the normal control by plasmin in the presence of [ethy-lenebis(oxyethylenenitrilo)]tetraacetic acid (EGTA).

Identifikasi Daging Tikus Pada Produk Baso Dengan Metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE)

2018 ◽  
Vol 26 (2) ◽  
pp. 058
Author(s):  
Anna P. Roswiem ◽  
Triayu Septiani
Keyword(s):  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Sds Page

<em>Bahan<strong> </strong>baku untuk membuat baso adalah daging hewan, pada umumnya dari daging sapi, ayam, ikan dan babi. Di beberapa daerah di Indonesia terjadi kasus baso tikus. Tujuan penelitian ini adalah menguji ada tidaknya kandungan daging tikus pada produk baso yang dijual di pasar Cempaka Putih-Kecamatan Kramat Jakarta Pusat dan di pedagang baso atau mie baso di sekitar kampus Universitas YARSI Jakarta. Daging adalah protein salah satu metode untuk mengidentifikasi protein adalah metode Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE).<strong> </strong>Hasil penelitian menunjukkan bahwa dari 6 sampel baso terindikasi ada 2 sampel baso dengan nomor 1 dan 5 yang dibuat dari campuran daging sapi dan tikus; ada 1 sampel baso dengan nomor 6 yang terbuat dari daging tikus; dan 2 sampel baso dengan nomor 2 dan 3 yang terbuat dari campuran sapi  dan babi, dan hanya 1 sampel baso dengan nomor sampel 4 yang benar-benar terbuat dari daging sapi.</em>

Majra Honey Abrogated the Normal and Cancer Cells Proliferation Inhibition by Juniperus procera Extract and Extract/Honey Generated AgNPs

2020 ◽  
Vol 20 (8) ◽  
pp. 970-981
Author(s):  
Hamed A. Ghramh ◽  
Essam H. Ibrahim ◽  
Mona Kilnay
Keyword(s):  
Anticancer Activity ◽  
Cancer Cells ◽  
Biological Activities ◽  
Sugar Content ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Bioactive Molecules ◽  
Sds Page ◽  
Splenic Cells ◽  
Juniperus Procera

Background: Juniperus procera and Majra honey are well-known as a folk medicine in many countries. Objectives: This work aimed to study the immunomodulatory effects after mixing Majra honey, J. procera water leaves extract and silver Nanoparticles (AgNPs) on immune or cancer cells. Methods: Juniperus procera water leaves extract and 20% Majra honey were prepared. Both the extract and honey were used separately to synthesize AgNPs. AgNPs were characterized using UV/Vis spectrophotometry and electron microscopy. Bioactive molecules in honey and the extract were explored using Fourier Transform Infrared (FT-IR) spectroscopy. Protein profile of honey was explored using Sodium Dodecyl Sulfate- Polyacrylamide Gel Electrophoresis (SDS-PAGE) and honey sugar content was determined using High- Performance Liquid Chromatography (HPLC). Biological activities of honey and the extract were tested. Results: The results demonstrated the ability of the extract/honey to produce AgNPs in a spherical shape. The extract/honey contained many functional groups. SDS-PAGE of Majra honey showed many protein bands. HPLC revealed honey is of good quality and no external additives are added to it. The extract and extract+ AgNPs inhibited the growth of normal rat splenic cells while honey stimulated it. The extract+honey turned stimulatory to the splenic cells’ growth and significantly diminished the inhibitory potential of the extract containing AgNPs. Both the extract and honey have antimicrobial activities, this potential increased in the presence of AgNPs. Honey and Honey+AgNPs inhibited HepG2 cancer cell proliferation while Hela cell growth inhibited only with honey+AgNPs. Conclusion: Both honey and the extract have antibacterial and immunomodulatory potentials as well as the power to produce AgNPs. Majra honey alone showed anticancer activity against HepGe2 cells, but not against Hela cells, and when contained AgNPs had anticancer activity on both cell lines. Mixing of Majra honey with J. procera extract showed characterized immunomodulatory potentials that can be described as immunostimulant.

scholarly journals Study on cocoonase, sericin, and degumming of silk cocoon: computational and experimental

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Preeti Anand ◽  
Jay Prakash Pandey ◽  
Dev Mani Pandey
Keyword(s):  
San Francisco ◽  
Tertiary Structure ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Evolutionary Genetics ◽  
Imaging Analysis ◽  
Silk Cocoon ◽  
Natural Color ◽  
Sds Page

Abstract Background Cocoonase is a proteolytic enzyme that helps in dissolving the silk cocoon shell and exit of silk moth. Chemicals like anhydrous Na2CO3, Marseille soap, soda, ethylene diamine and tartaric acid-based degumming of silk cocoon shell have been in practice. During this process, solubility of sericin protein increased resulting in the release of sericin from the fibroin protein of the silk. However, this process diminishes natural color and softness of the silk. Cocoonase enzyme digests the sericin protein of silk at the anterior portion of the cocoon without disturbing the silk fibroin. However, no thorough characterization of cocoonase and sericin protein as well as imaging analysis of chemical- and enzyme-treated silk sheets has been carried out so far. Therefore, present study aimed for detailed characterization of cocoonase and sericin proteins, phylogenetic analysis, secondary and tertiary structure prediction, and computational validation as well as their interaction with other proteins. Further, identification of tasar silkworm (Antheraea mylitta) pupa stage for cocoonase collection, its purification and effect on silk sheet degumming, scanning electron microscope (SEM)-based comparison of chemical- and enzyme-treated cocoon sheets, and its optical coherence tomography (OCT)-based imaging analysis have been investigated. Various computational tools like Molecular Evolutionary Genetics Analysis (MEGA) X and Figtree, Iterative Threading Assembly Refinement (I-TASSER), self-optimized predicted method with alignment (SOPMA), PROCHECK, University of California, San Francisco (UCSF) Chimera, and Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) were used for characterization of cocoonase and sericin proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), protein purification using Sephadex G 25-column, degumming of cocoon sheet using cocoonase enzyme and chemical Na2CO3, and SEM and OCT analysis of degummed cocoon sheet were performed. Results Predicted normalized B-factors of cocoonase and sericin with respect to α and β regions showed that these regions are structurally more stable in cocoonase while less stable in sericin. Conserved domain analysis revealed that B. mori cocoonase contains a trypsin-like serine protease with active site range 45 to 180 query sequences while substrate binding site from 175 to 200 query sequences. SDS-PAGE analysis of cocoonase indicated its molecular weight of 25–26 kDa. Na2CO3 treatment showed more degumming effect (i.e., cocoon sheet weight loss) as compared to degumming with cocoonase. However, cocoonase-treated silk cocoon sheet holds the natural color of tasar silk, smoothness, and luster compared with the cocoon sheet treated with Na2CO3. SEM-based analysis showed the noticeable variation on the surface of silk fiber treated with cocoonase and Na2CO3. OCT analysis also exemplified the variations in the cross-sectional view of the cocoonase and Na2CO3-treated silk sheets. Conclusions Present study enlightens on the detailed characteristics of cocoonase and sericin proteins, comparative degumming activity, and image analysis of cocoonase enzyme and Na2CO3 chemical-treated silk sheets. Obtained findings illustrated about use of cocoonase enzyme in the degumming of silk cocoon at larger scale that will be a boon to the silk industry.

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