Evaluation of effective protein extraction procedure to profile petiole of Moringa oleifera

Moringa oleifera is widely known as multipurpose tree since all of its parts confer multiple functions. The leaf is highly favourable among consumers while the petiole is mostly wasted. There are numerous studies on the flavonoid and antioxidant property of the stem and twig. However, study on the petiole has never been done. There-upon, this study was conducted to develop protein profiling of the petiole. In this study, 6 different protein extraction methods were tested on the fresh petiole before its protein quantity and quality were checked via Bradford assay and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) respectively. The in-solution digestion was then done prior to LC-MS/MS analysis. The protein electrophoretic pattern from the SDS-PAGE proves that method 6 using Tris HCl buffer with incorporation of dithiothreitol (DTT) and phenylmethylsulfonyl fluoride (PMSF) confers the best quality of protein. It produced the highest number of visible individual bands compared to other methods. Meanwhile, 93 proteins were successfully identified via LCMS analysis where the protein, signal response and carbohydrate metabolism categories confer the highest percentage. High quality and content of the protein extracted from the petiole including the antioxidant, anticancer and antidiabetic protein identified suggested that consuming this part of the plant could enhance nutrients of human body.

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Detección inmunoquímica de la adulteración de chorizo de cerdo con proteínas de soja

Food Science and Technology International ◽

10.1177/108201329800400404 ◽

1998 ◽

Vol 4 (4) ◽

pp. 257-262 ◽

Cited By ~ 18

Author(s):

A.F. González-Córdova ◽

A.M. Calderón de la Barca ◽

M. Cota ◽

B. Vallejo-Córdoba

Keyword(s):

Gel Electrophoresis ◽

Protein Extraction ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Electrophoretic Pattern ◽

Soy Flour ◽

Nacl Solution ◽

Standard Curve ◽

Sds Page ◽

Polyclonal Antisera

Rabbit polyclonal antisera were produced against soy flour proteins extracted with 0.5 M NaCl solution and purified by affinity chromatography to detect adulteration in pork chorizo (sausage) with an enzyme-linked immunoabsorbent assay (ELISA). To detect adulteration the following saline extracts were prepared: 100% pork and 100% soy chorizo and mixtures of these two pro ducts (90:10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90), and extracts of two commercial chorizos labeled as pork. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the salt solution was the most suitable for protein extraction compared to water and a 1% SDS solution. The commercial chorizos did not have the characteristic electrophoretic pattern for pork chorizos. The antibodies were specific for soy and its products, as there was no response to other vegetable and animal proteins by gel immunodiffusion assay or ELISA. The standard curve obtained with the ELISA results of the chorizo mixtures gave a correlation coefficient of r2 = 0.988. The two commercial chorizo values resulted in a 32 and 40% soy substitution. Total time for ELISA was optimized to 4 h, and 2 h if the plates were previously activated with the antigen. Variation coefficients of ELISA readings for replicates of the same extract in one plate and variation of different assays on different days were 2.3 and 8% respectively.

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Assessment of Some Bacteria from Panteka Stream, Kaduna, Nigeria, for their Larvicidal Activity Against Anopheles gambiae

European Journal of Engineering Research and Science ◽

10.24018/ejers.2018.3.12.1025 ◽

2019 ◽

Vol 3 (12) ◽

pp. 149-154

Author(s):

Thankgod Ositadinma Ndibe ◽

Nancy Erika Nwabufo ◽

Johnson John Usman ◽

Winnie Chuno Eugene

Keyword(s):

Anopheles Gambiae ◽

Larvicidal Activity ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Protein Profiling ◽

Mosquito Larvae ◽

Bacterial Isolates ◽

Sample Collection ◽

Sds Page ◽

Biochemical Identification

It is obvious that malaria is one of the commonest diseases in Africa, hence the need to embark on a study to reduce its transmission by eliminating the vector. Some microorganisms are known to have larvicidal activity leading to destruction of mosquito larvae, thereby, preventing them from metamorphosing into adult mosquitoes that can transmit Plasmodium spp. Panteka stream, Kaduna, Nigeria, is a dumping site for refuse and automobile waste and thus, a potential source of bacteria. This present investigation was aimed at screening bacterial isolates for their larvicidal activity against Anopheles gambiae. Standard methods were employed in sample collection, isolation, morphological, biochemical identification and protein profiling of these bacteria isolates. Five different types of bacteria were identified; Bacillus thuringiensis, Staphylococcus aureus, Micrococcus sedentarius, Enterococcus faecalis and Streptococcus pneumonia. Among these bacteria, B. thuringiensis exhibited the most larvicidal activity, followed by M. sedentarius. On the basis of lethal concentration (LC50), B. thuringiensis exhibited the highest lethal activity against Anopheles gambiae larvae at 48 hour duration of exposure. Results showed that concentration of bacterial isolates and duration of exposure of larvae to the bacterial isolates, determine the mortality rate of larvae. Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) revealed variable bands between B. thuringiensis and M. sedentarius, which might have accounted for their differences in larvicidal activity. The use of bacteria for the control of mosquito larvae is highly recommended. Further research should be conducted to search for more bacteria and possibly fungi which have potentials for larvicidal activity.

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Evaluation of Genetic Diversity in Barley (Hordeum vulgare) Genotypes using Protein Profiling

Journal of Plant Breeding and Genetics ◽

10.33687/pbg.006.01.2449 ◽

2018 ◽

Vol 6 (1) ◽

pp. 23-31

Author(s):

Manal Eid

Keyword(s):

Genetic Diversity ◽

Hordeum Vulgare ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Protein Profiling ◽

Protein Patterns ◽

Systematic Classification ◽

Molecular Weights ◽

Sds Page ◽

Solid Foundation

The knowledge of the genetic diversity of barley (Hordeum vulgare) genotypes based on protein polymorphism is very important for breeding programs. The purpose of the current study was to determine the genetic diversity and relationships among ten barley genotypes by using Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) for protein profiles. A total number of 30 bands with molecular weights ranging from 12 to 148 KD were detected. Out of these, five bands were observed monomorphic. Rest of the bands had shown polymorphism to the extent of 83.3% among the test genotypes. The genetic similarity of the ten genotypes tested varied from 0.26 to 1.00 with an average of 0.51. Cluster analysis divided the ten genotypes into two major clusters comprising four subclusters, which was consistent with the systematic classification of barley done in previous studies. The results of this study indicated that the genotypes of barley could effectively be differentiated based on polymorphism, detected between protein patterns. SDS-PAGE presented a higher differentiation power and better repeatability; thus, could be used as a rapid and reliable method for genetic diversity analysis and laid a solid foundation for future barley breeding.

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Revealing contrasting genetic variation and study of genetic diversity in urdbean (Vigna mungo (L.) Hepper) using SDS-PAGE of seed storage proteins

Research in Biotechnology ◽

10.19071/rib.2016.v7.2863 ◽

2016 ◽

Vol 7 ◽

Cited By ~ 1

Author(s):

Swapan Kumar Tripathy ◽

Priyadarshini Mohanty ◽

Monalisha Jena ◽

Gokul Bihari Dash ◽

Kedareswar Pradhan ◽

...

Keyword(s):

Genetic Variation ◽

Storage Protein ◽

Storage Proteins ◽

Seed Storage Protein ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Similarity Index ◽

Seed Storage ◽

Protein Profiling ◽

Sds Page

Total seed storage protein profiles of 20 urdbean genotypes including the popular variety T9 were analysed by Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). 14 genotypes could be clearly identified based on genotype-specific seed protein fingerprints while rest of the test genotypes were categorized into three protein types. Dendrogram based on electrophoretic data clustered the genotypes into seven groups at 78.5% phenon level. TU 95-1 with TU 12-25-4 revealed lowest similarity index value (0.33) followed by TU 95-1 with PU 30 and KU 96-3(SI=0.35). Clustering pattern revealed distinctly divergent group formed by TPU 95-1 and TPU 4. These may serve as a valuable source genotype in recombination breeding. Key words: Seed storage protein profiling, SDS-PAGE, Genetic variation, urdbean.

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Crystalline protein profiling and cry gene detection in Bacillus thuringiensis strains isolated during epizootics in Cydia pomonella L.

Biological Letters ◽

10.1515/biolet-2015-0008 ◽

2014 ◽

Vol 51 (2) ◽

pp. 83-92 ◽

Cited By ~ 2

Author(s):

Edyta Konecka ◽

Jakub Baranek ◽

Adam Kaznowski

Keyword(s):

Bacillus Thuringiensis ◽

Polyacrylamide Gel Electrophoresis ◽

Cydia Pomonella ◽

Sodium Dodecyl ◽

Natural Populations ◽

Protein Profiling ◽

Gene Detection ◽

Sds Page ◽

Molecular Masses ◽

Reference Strains

AbstractThe composition of Bacillus thuringiensis crystalline inclusions was characterized in 18 strains: 12 isolates were obtained from the intestinal tract of Cydia pomonella larvae during epizootics, 2 isolates were cultured from Leucoma salicis larvae taken from their natural populations, and 4 reference strains. The number and molecular mass of B. thuringiensis crystalline proteins (Cry and Cyt) was estimated by the sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The crystals contained 1-8 proteins with molecular masses of 36-155 kDa. The toxin profiles differed both quantatively and qualitatively. The B. thuringiensis MPU B9 isolate had the highest number and diversity of Cry toxins. The analysis of crystal composition by SDS-PAGE was insufficient to detect groups and subgroups of Cry proteins. We identified 20 groups and 3 subgroups of Cry and Cyt crystalline toxins. Only one epizootic strain harboured cry25. In single reference strains, the cry1H, cry10 and cry25 genes were found. We did not find any correlation between the occurrence of cry genes and electrophoretic protein profiles of crystalline toxins.

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A Proteomic Approach to Identify Zein Proteins upon Eco-Friendly Ultrasound-Based Extraction

Biomolecules ◽

10.3390/biom11121838 ◽

2021 ◽

Vol 11 (12) ◽

pp. 1838

Author(s):

Laura Darie-Ion ◽

Madhuri Jayathirtha ◽

Gabriela Elena Hitruc ◽

Marius-Mihai Zaharia ◽

Robert Vasile Gradinaru ◽

...

Keyword(s):

Protein Extraction ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Protein Composition ◽

Industrial Applications ◽

Experimental Conditions ◽

Proteomic Approach ◽

Sds Page ◽

Pharmaceutical Industries

Zein is a type of prolamin storage protein that has a variety of biomedical and industrial applications. Due to the considerable genetic variability and polyploidity of the starting material, as well as the extraction methods used, the characterization of the protein composition of zein requires a combination of different analytical processes. Therefore, we combined modern analytical methods such as mass spectrometry (MS), Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), atomic force microscopy (AFM), or Fourier transform infrared spectroscopy–attenuated total reflectance (FTIR-ATR) for a better characterization of the extracted zein. In this study, we present an enhanced eco-friendly extraction method, including grinding and sieving corn seeds, for prolamins proteins using an ultrasonic extraction methodology. The use of an ultrasonic homogenizer, 65% ethanol extraction buffer, and 710 µm maize granulation yielded the highest protein extraction from all experimental conditions we employed. An SDS PAGE analysis of the extracted zein protein mainly revealed two intense bands of approximatively 20 and 23 kDa, suggesting that the extracted zein was mostly α-zein monomer. Additionally, MS analysis revealed as a main component the α-zein PMS2 (Uniprot accession no. P24450) type protein in the maize flour extract. Moreover, AFM studies show that extracting zein with a 65% ethanol and a 710 µm granulation yields a homogeneous content that could allow these proteins to be employed in future medical applications. This research leads to a better understanding of zeins content critical for developing new applications of zein in food and pharmaceutical industries, such as biocompatible medical vehicles based on polyplexes complex nanoparticles of zein with antimicrobial or drug delivery properties.

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

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646313 ◽

1992 ◽

Vol 68 (05) ◽

pp. 534-538 ◽

Cited By ~ 5

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).

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

YARSI Medical Journal ◽

10.33476/jky.v26i2.394 ◽

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

Bahan 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). 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.

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

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200213104224 ◽

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.

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A comparative study on the extraction effects of common agents on collagen-based binders in mural paintings

Heritage Science ◽

10.1186/s40494-021-00519-y ◽

2021 ◽

Vol 9 (1) ◽

Author(s):

Jianghao Du ◽

Zhanyun Zhu ◽

Junchang Yang ◽

Jia Wang ◽

Xiaotong Jiang

Keyword(s):

Protein Structure ◽

Comparative Study ◽

Protein Extraction ◽

Guanidine Hydrochloride ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Extraction Process ◽

Ultraviolet Absorption Spectrum ◽

Mural Paintings ◽

The Impact

AbstractIn this paper, a comparative study was conducted on the extraction effects of six agents for collagen-based mural painting binders. These agents were used to extract the residual proteins in the non-aged and thermal aged samples. The protein extraction efficiencies of different extracting agents were quantitatively determined by bicinchoninic acid (BCA) method, and then processed by multivariate analysis of variance (MANOVA). The impact of the extraction process on the protein structure was characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), ultraviolet absorption spectrum (UV) and circular dichroism (CD). The results showed that, for both non-aged and aged samples, the extraction efficiency of 2 M guanidine hydrochloride (GuHCl) was significantly higher than the other five agents, with less damage to the protein structure during the extraction process.

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