PROTEIN PROFILING OF DIFFERENT PLANT TISSUES FROM HERB PHYLLANTHUS NIRURI

2015 ◽  
Vol 77 (24) ◽  
Author(s):  
Ainul Mardhiah Mohd Nail ◽  
Noor Hasniza Md Zin
Keyword(s):  
Molecular Weight ◽  
Protein Band ◽  
Protein Profiling ◽  
Protein Extract ◽  
Phyllanthus Niruri ◽  
Two Dimensional Gel Electrophoresis ◽  
Plant Parts ◽  
Sds Page ◽  
Isoelectric Points

Herb Phyllanthus niruri (P. niruri) is known to have various pharmacological functions including anticancer, antibacterial, antioxidant, anti-hypertensive and also anti-diabetic properties.  In this research, the proteomic part of P. niruri was studied to determine the bioactive peptides that responsible for specific characteristics. Total soluble proteins from different plant parts of freshly collected P. niruri were extracted using TCA/acetone method and then quantified using Bradford assay. Fruits part was found to have a significantly higher amount of proteins (4.91µg/µl + 0.21) compared to leaves (4.18µg/µl + 0.15). To determine the quality of proteins in the crude extract, SDS-Page was carried out which separates proteins in the basis of molecular weight. Proteins extracted from leaves were widely distributed between the range of 3.5 kDa to 160 kDA. Meanwhile, proteins in fruits mainly distributed within the range of 15 kDa to 80 kDa. The most highly expressed protein band was found in fruit, located in between 30 to 40 kDa. The protein extracts were then further analyzed based on the molecular weight and isoelectric points using two-dimensional gel electrophoresis (2D-GE) approach. Based on the profile pattern obtained from 2D-GE analysis, protein extract from fruits seems to express more protein spots compared to protein extract from leaves. Protein spots from fruit are seen to be intensely resolved within pH 4 to 10 at molecular weight between 10 kDa to 80 kDa. On the other hand, protein spots from leaves were moderately resolved at pH 4 to 10 at molecular weight within 10 kDa to 50 kDa.

scholarly journals Protein profiling of chicken breast muscles from different slaughter houses in relation to meat quality

2020 ◽  
Vol 15 (1) ◽  
Author(s):  
Noor Hasniza Zin ◽  
Murni Abdul Halim ◽  
Noraslinda Muhamad Bunnori ◽  
Normah Haron ◽  
Widya Abdul Wahab
Keyword(s):  
Molecular Size ◽  
Protein Band ◽  
Poor Quality ◽  
Vital Role ◽  
Chicken Meat ◽  
Protein Profiling ◽  
Chicken Breast ◽  
Protein Profiles ◽  
Αb Crystallin

Introduction: Chicken meat is a source of protein in the human diet. Protein content and values define the quality of chicken meat. This research aimed to analyze variations of protein profiles in chicken breast muscles from different slaughtering houses by using proteomic strategies. Methods: Total proteins of chicken breast muscles from three different slaughtering houses (Sample A, Sample B and Sample C) were extracted and quantified by using Bradford assay. Then, the proteins were separated by SDS-PAGE to monitor the quality of extracted proteins. Protein profiles in different samples were compared by 2D-GE analysis. Results: The most highly expressed protein band was located between the molecular size of 37-50 kD in all samples and it was expected to be betaactin. While resolved in 2D-PAGE, differences in protein expression were observed between samples. There were three spots expressed with highest intensity in Sample B compared to others. The protein spot detected at pH 5.28 and the size range between 50- 75 kD was predicted to be NADPH-cytochrome P450 reductase (CPR), at pH 6.45 and molecular weight between 37-50 kD was expected to be creatine kinase M-type (M-CK) while at pH 6.78 and molecular size nearly 25 kD was expected to be αB-crystallin. Conclusions: It can be mentioned that these proteins could play a vital role in mechanisms that contribute to the poor quality of chicken meat.

scholarly journals Variations in the Methionine-rich Protein Composition of the Genus Arachis1

1984 ◽  
Vol 11 (1) ◽  
pp. 1-3 ◽  
Author(s):  
Sheikh M. Basha ◽  
Sunil K. Pancholy
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Protein Composition ◽  
Electrophoretic Analysis ◽  
Two Dimensional ◽  
Weight Group ◽  
Two Dimensional Gel Electrophoresis ◽  
One Dimensional ◽  
Molecular Weights ◽  
Isoelectric Points

Abstract Methionine-rich proteins (MRP) from seeds of different species of the Genus Arachis were isolated and analyzed by gel electrophoresis to detect possible compositional differences. One-dimensional gel electrophoretic analysis showed presence of quantitative and qualitative variations among the MRP-fractions. Following two-dimensional gel electrophoresis, the MRP-fractions were found to contain three groups of polypeptides with apparent molecular weights of approximately 21,000; 19,000 and 16,000, and isoelectric points between 5.1 and 5.8. Within each molecular weight group the number of polypeptides varied between 1 and 3.

scholarly journals The Potential of Candida Biofilm Protein as Bioreceptor for Candidiasis Immunoassay

2018 ◽  
Vol 14 (1) ◽  
pp. 47 ◽  
Author(s):  
Masfufatun Masfufatun ◽  
Loo Haryanto ◽  
Harsono Harsono
Keyword(s):  
Molecular Weight ◽  
Candida Albicans ◽  
Polyclonal Antibody ◽  
Control Group ◽  
Potential Candidate ◽  
Dot Blot ◽  
Protein Extract ◽  
Treatment Groups ◽  
Sds Page ◽  
Mechanical Methods

Abstract: Candidiasis or infection that is caused by Candida has become a new list of the therapeutical problems recently. The difficulties in diagnosing are the main cause of the unsatisfactory results from common therapies and diagnosis methods. This has urged researchers to find alternative ways in candidiasis diagnosis such as serology-based detection using antigen or antibody development. The aim of this study was to evaluate the potential of protein derived from Candida albicans biofilm as bioreceptor on candidiasis immunoassay through Dot Blot method. The research method used descriptive method with the following stages: (1) preparation of Candida albicans biofilm (2) extraction of Candida albicans protein through enzymatic and mechanical methods, (3) determination of protein molecular weight with SDS-PAGE (4) production of polyclonal anti- candida and (5) analysis of protein extract as bioreeceptor on dot blot. Profile of biofilm proteins on SDS-PAGE analysis were shown on molecular weight 27,42; 29,89; 38,10; 44,90; 48,75; 52,92; 55,14; 59,86; 70,56; 87,36; 102,54;115,05; 130,14;143,14;181,53 kD. There were differences in the intensity of dots in the control group (44070) and treatment groups (63170.5). It is noticeable that biofilm protein extract of C. albicans can be used for induction of anti-Candida polyclonal antibody production as the potential candidate of bioreceptor in candidiasis immunoassay. Keywords: SDS-PAGE, polyclonal antibody, immunoassay, dot blot, biofilm

scholarly journals Characterization of Inhibin from Culture and Non Culture of Granulose Cells for Monoclonal Antibody of Inhibin Production

2010 ◽  
Vol 4 (1) ◽  
Author(s):  
Amiruddin Amiruddin ◽  
Tongku Nizwan Siregar ◽  
Amalia Sutriana ◽  
Dwinna Aliza ◽  
T. Armansyah
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Western Blot ◽  
Total Amino Acid ◽  
Multiple Ovulation ◽  
Sds Page ◽  
Isoelectric Points ◽  
Granulose Cells

This study has long-term objectives to obtain immunogenic prototype that can be used to induce multiple ovulation in goats. Working steps of this study were begun with the collection of ovarium from goats, collection of granulose cells, culture of granulose and characterization of molecular weight and isoelectric point (pI) of inhibin protein of granulose cells obtained from culture and non-culture of granulose cells, and followed by preparation of monoclonal antibody toward inhibin. The results showed that inhibin isolated either from culture or non-culture of granulose cells produced a 32 kDa band. Molecular weight of inhibin was measured by Western Blot. The 32 kDa band of SDS PAGE product appeared on Western Blot result was inhibin molecules produced by granulose cells collected fom culture and non-culture of granulose cells that can be identified by Mab-inhibin. Product of IEF gel electrophoresis suggested that inhibin molecule collected from culture of granulose cells has no charge at isoelectric points ranging from 5-6, depends on its total amino acid composition.

scholarly journals Grain quality of common wheat lines with T. kiharae genetic material

2018 ◽  
Vol 23 ◽  
pp. 108-113
Author(s):  
O. A. Orlovskaya ◽  
S. I. Vakula ◽  
L. V. Khotyleva ◽  
A. V. Kilchevsky
Keyword(s):  
Molecular Weight ◽  
Common Wheat ◽  
Grain Quality ◽  
Genetic Material ◽  
Glutenin Subunits ◽  
Gluten Quality ◽  
Gluten Content ◽  
Sds Page ◽  
Wheat Lines

Aim. T. kiharae (AtAtGGDD, 2n=42) is a source of high protein and gluten content, resistance to many diseases. Сommon wheat lines with the introgression of T. kiharae genetic material were obtained in order to enrich T. aestivum L. gene pool. The aim of this study was to assess the impact of T. kiharae genetic material on the grain quality of T. aestivum/T. kiharae introgression lines. Methods. The composition of the high molecular weight glutenin subunits was analyzed by SDS-PAGE. Evaluation of the most important traits of grain quality (hardness, protein and gluten content, gluten quality) was carried out according to GOST. Results. Сomparative analysis of the composition of high molecular weight glutenin subunits of introgressive lines and their parental forms allowed us to identify lines with novel alleles of Glu-1 loci, specific for T. kiharae. For most of the introgression lines T. aestivum/T. kiharae hardness, protein and gluten content were higher than for parent wheat varieties. Conclusions. Introgression of T. kiharaegenetic material in the genome of common wheat had a positive effect on all studied parameters of grain quality except the gluten quality. Keywords: common wheat, T. kiharae, glutenin, SDS-PAGE, quality of grain.

scholarly journals Quality of Cultured Wader Pari During Storage at Different Temperature

2017 ◽  
Vol 20 (2) ◽  
pp. 339 ◽  
Author(s):  
Mala Nurilmala ◽  
Agoes Mardiono Jacoeb ◽  
Rofi Ahmad Dzaky
Keyword(s):  
Molecular Weight ◽  
Functional Group ◽  
Gel Strength ◽  
Yellowfin Tuna ◽  
Fish Skin ◽  
Alternative Source ◽  
Food Industries ◽  
Sds Page ◽  
Fish Skin Gelatin

Gelatin is one of the products which become a necessity for various industries, i.e. food and non-food industries. The application of gelatin has been increasing year by year in Indonesia. However, there is no<br />gelatin industry in Indonesia so far. Thus, it is necessary to find an alternative source of gelatin, especially from fishery by products.Thus, the purpose of this research was to extract fish skin gelatin of yellowfin tuna with temperature treatments (55, 65 and 75oC). In addition, the properties of resulted gelatin were determined including yield, pH, gel strength, viscosity, functional groups, molecular weight profiles, and amino acid composition. The extraction at 75oC was chosen as the best result. The yield was 17%; pH 5.3; gel strength 1789.55 gf, viscosity 104.2 Cp, respectively. There was functional group amide A, I, II, dan III. SDS-PAGE showed β, α1 dan α2 bands for tuna skin gelatin. In addition, the main amino acids were glycine and proline.

scholarly journals Protein Expression of Soybean (Glycine Max L. Merr) Varieties In Drought Stress

el–Hayah ◽  
2017 ◽  
Vol 6 (2) ◽  
pp. 50
Author(s):  
Evika Sandi Savitri ◽  
Estri Laras Arumingtyas
Keyword(s):  
Drought Stress ◽  
Protein Expression ◽  
Molecular Study ◽  
Protein Band ◽  
Drought Condition ◽  
Two Dimensional Gel Electrophoresis ◽  
Sds Page ◽  
Stress Changes ◽  
Glycine Max L ◽  
New Protein

<em>Drought is one of the most severe limitations on the productivity of soybean. There are many genes and proteins involved in drought stress tolerance.  Identification of proteins which could be used as the base for the development of molecular study is very important to understand drought tolerance thoroughly. The objective of the research was to investigate protein expression of soybean to drought stress. Changes in protein expression were analyzed using SDS PAGE and two dimensional gel electrophoresis. Image analysis of 2D protein was performed by using the PDQuest 8.0 software program (Bio-Rad). Tolerant variety, Dering-1, was subjected to drought stress using limitation of watering, while Detam-1, a sensitive variety, was used as comparator.  The result showed that protein concentration have decreased in drought condition from 3,22 mg/ml to 0,77 mg/ml. The new protein band with the 24,95 kDa have been found in drought condition. This protein is osmotin like protein with the accession number NP915414 which may play a role in the mechanism of drought resistance. The identification of the protein based on sequence amino acid literature review</em>

scholarly journals Prekeratin biosynthesis in human scalp epidermis

1982 ◽  
Vol 208 (1) ◽  
pp. 179-187 ◽  
Author(s):  
P T Bladon ◽  
P E Bowden ◽  
W J Cunliffe ◽  
E J Wood
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl ◽  
Post Translational Modification ◽  
Two Dimensional Gel Electrophoresis ◽  
Keratin Gene ◽  
One Dimensional ◽  
Electrophoretic Mobilities ◽  
Isoelectric Points ◽  
Intense Labelling ◽  
Dimensional Electrophoresis

Analysis of human scalp epidermal prekeratin polypeptides by two-dimensional gel electrophoresis revealed that each of the bands observed in one-dimensional electrophoresis consisted of three to five polypeptides of the same molecular weight but differing in isoelectric points. It was possible to divide the polypeptides into two families, with isoelectric points in the ranges pH 6.0-8.0 and pH 5.0-5.5 respectively. Incorporation of radiolabelled amino acids into freshly excised pieces of scalp epidermis showed that some of the polypeptides had relatively greater contents of glycine and serine than others. Radiolabelled methionine and leucine were, in contrast, incorporated more or less uniformly into all the polypeptides. After incubation with 32P-labelled orthophosphate, relatively more intense labelling by 32P was observed in the higher molecular weight bands of each family. The most basic of the isoelectric variants in each case did not take up phosphate, implying that at least some of the variation in charge was due to different degrees of phosphorylation. Polyadenylated RNA isolated from scalp epidermis was translated in an RNA-dependent reticulocyte haemolysate system followed by immunoprecipitation and electrophoresis. The polypeptides isolated by using anti-(human scalp prekeratin) immunoglobulin G had similar electrophoretic mobilities in sodium dodecyl sulphate/polyacrylamide gels to authentic prekeratin polypeptides, but had different isoelectric properties. This suggested that the products of keratin gene expression undergo post-translational modification.

scholarly journals Characterization and Identification of Allergen Protein in Shrimp Before and After Heated with SDS-PAGE Method

2019 ◽  
Vol 2 (2) ◽  
pp. 87-93
Author(s):  
Emma Emawati ◽  
Idar Idar ◽  
Resta Ramadiyanti
Keyword(s):  
Molecular Weight ◽  
Anion Exchange ◽  
Column Chromatography ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Band ◽  
Food Allergies ◽  
Chromatography Method ◽  
Anion Exchange Column ◽  
Sds Page

Food allergies are one of the most common allergies in Indonesian society. Generally, when children aged 5-6 years food allergies will disappear, except peanut allergies and allergies to seafood, such as fish, shellfish and crustaceans. This study aims to determine the pattern of separation of allergen proteins in shrimp using anion exchange column chromatography method and identify allergen proteins in shrimp using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) method. Protein extraction from shrimp using Phosphate buffer saline (PBS) pH 7.2 and centrifuged at 10,000 rpm for 10 min at 4�C. Protein separation was carried out by anion-exchange column chromatography method, and the fraction obtained was measured at 280nm wavelength. The highest yield at absorbance was identified by using SDS-PAGE. Polyacrylamide gel electrophoresis was used to determine the protein profile and molecular weight of shrimp extract. Coloring of protein bands using silver staining. Data were analyzed descriptively based on the migration value of the sample protein bands compared to the marker protein band (Rf). The results of protein allergen profile analysis on shrimp using SDS-PAGE showed that the shrimp contained a protein band with a molecular weight of 37.77 kDa for cooked shrimp and 37.03 kDa for fresh shrimp.

scholarly journals Polypeptide Composition of Arachin and Non-arachin Proteins From Early Bunch Peanut (Arachis hypogaea L.) Seed1

1981 ◽  
Vol 8 (2) ◽  
pp. 82-88 ◽  
Author(s):  
Shaik-M. M. Basha ◽  
Sunil K. Pancholy
Keyword(s):  
Molecular Weight ◽  
Arachis Hypogaea ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Seed Proteins ◽  
Two Dimensional ◽  
Polypeptide Composition ◽  
Two Dimensional Gel Electrophoresis ◽  
Arachis Hypogaea L ◽  
Isoelectric Points

Abstract Peanut (Arachis hypogaea L.) seed proteins were resolved into arachin and non-arachin fractions, and composite two-dimensional polypeptide maps were prepared. Seed proteins were extracted with a buffer containing 2 M NaCl, 10 mM Tris-HCl (pH 8.2), 0.2 mM phenylmethyl sulfonyl fluoride and 0.002% NaN3 and resolved into ten peaks by gel filtration on a Sephacryl S-300 column. Gel filtration of total protein extract yielded three molecular weight variants (490,000., 400,000, and 365,000) of arachin. Gel electrophoresis showed quantitative and qualitative differences in the protein and polypeptide composition of the three arachin variants. Nonarachin proteins obtained by this method were heterogeneous and distinct from the arachin. Two-dimensional gel electrophoresis revealed several differences in the polypeptide composition between arachin fraction IV and fractions II and III. Composite two-dimensional polypeptide maps of arachin and non-arachin revealed the presence of several polypeptides with similar isoelectric points and molecular weights between them. Arachin contained six molecular weight (between 15,500 and 68,000) classes of polypeptides with isoelectric points between 4.7 and 8.4 while nonarachin contained nine molecular weight (between 16,000 and 170,000) classes of polypeptides having isoelectric points between 4.7 and 7.9.

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