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Search Results: 1 - 10 of 1005 matches for " sds-page "
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Purification and characterization of the kinetic parameters of cellulase produced from wheat straw by Trichoderma viride under SSF and its detergent compatibility  [PDF]
Hafiz Muhammad Nasir Iqbal, Ishtiaq Ahmed, Muhammad Anjum Zia, Muhammad Irfan
Advances in Bioscience and Biotechnology (ABB) , 2011, DOI: 10.4236/abb.2011.23024
Abstract: This paper reports the purification and characterization of kinetic parameters of cellulase produced from Trichoderma viride under still culture solid state fermentation technique using cheap and an easily available agricultural waste material, wheat straw as growth supported substrate. Trichoderma viride was cultured in fermentation medium of wheat straw under some previously optimized growth conditions and maximum activity of 398±2.43U/mL obtained after stipulated fermentation time period. Cellulase was purified 2.33 fold with specific activity of 105U/mg in comparison to crude enzyme extract using ammonium sulfate precipitation, dialysis and Sephadex-G-100 column chromatography. The enzyme was shown to have a relative low molecular weight of 58kDa by sodium dodecyl sulphate poly-acrylamide gel electrophoresis. The purified enzyme displayed 6.5 and 55oC as an optimum pH and temperature respectively. Using carboxymethyl cellulose as substrate, the enzyme showed maximum activity (Vmax) of 148U/mL with its corresponding KM value of 68µM. Among activators/inhibitors SDS, EDTA, and Hg2+ showed inhibitory effect on purified cellulase whereas, the enzyme activated by Co2+ and Mn2+ at a concentration of 1mM. The purified cellulase was compatible with four local detergent brands with up to 20 days of shelf life at room temperature suggesting its potential as a detergent additive for improved washing therefore, it is concluded that it may be potentially useful for industrial purposes especially for detergent and laundry industry.
Characterization of Aeromonas hydrophila through RAPD-PCR and SDS-PAGE Analysis  [PDF]
Agniswar Sarkar, Mousumi Saha, Avijit Patra, Pranab Roy
Open Journal of Medical Microbiology (OJMM) , 2012, DOI: 10.4236/ojmm.2012.22005
Abstract: Aeromonas hydrophila isolated from fish (Labeo rohita), pond water, river water, raw meat of chicken and mutton and raw cow milk were characterized through Random Amplified Polymorphic DNA (RAPD) analysis and Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS PAGE) of cellular proteins. RAPD-PCR amplification of bacterial DNA was done by using ten random primers (OPA-01 to 10) and found some distinct banding pattern on agarose gel. RAPD profile was studied with each isolate and absolute polymorphism indicating its application as an ideal tool for molecular characterization. Other methods like morphological, serological and biochemical characterization gives contradictory results and total cellular protein profiling does not show any significant polymorphism for identification and discrimination.
Diversity analysis of chickpea (Cicer arietinum L.) germplasm and its implications for conservation and crop breeding  [PDF]
Zakia Ahmad, Abdul Samad Mumtaz, Mohammad Nisar, Nasrullah Khan
Agricultural Sciences (AS) , 2012, DOI: 10.4236/as.2012.35087
Abstract: The exploration of genetically variable accessions is the key source of germplasm conservation and potential breeding material for the future. The more diverse group of cultivars provides an ample opportunity to breeders for releasing new and superior varieties, considering their quality traits for direct commercial utilization. In this study, we assessed the genetic diversity of Cicer arietinum 70 accessions from Pakistan and USA using morphological traits, seed protein and molecular markers. Based on four morphological traits, the average coefficient of variation was calculated as 56.8% with significant correlation among yield traits. The analysis revealed that the accessions 1898, 2819, 3022, 3037, 3040, 3043, 3054, 3059 and 3063 were best in performance with a total of 12% environmental error. The cluster analysis based on protein data revealed 50% genetic diversity among accessions. The clustering pattern did not show any grouping that could be attributed to either the geographic distribution or the field performance. For molecular characterization of germplasm 5 PCR based RAPD primers, OPA4, OPA9, OPG13, UBC181 and UBC733b used were found to be polymorphic with 37% genetic diversity among local and exotic accessions. Whereas, 3 SSR primers viz., CaSTMS2, Ca- STMS15 and CaSTMS21 scored the genetic variability up to 55% by cluster analysis through UPGMA percent disagreement. The primers, TA72 and TA130 were linked with yield related traits, indicated highest dissimilarity index value (0.69) and notable variation in the identified promising lines. The Morphometric, Biochemical and Molecular markers reported here, are helpful to assess the extent of genetic diversity among Chickpea accessions and can be used to identify the unreported cultivars with desirable quantitative traits for improving Chickpea yield in Pakistan. Based on the study, the accessions 3043 and 3054 have been recommended to the breeders for their future use in multiplication and screening against various diseases.
Digitonin and sodium dodecylsulfate-solubilized frog rho-dopsin: Behavior under native and denaturing polyacrylamide gel electrophoresis  [PDF]
Sergey A. Shukolyukov
Advances in Biological Chemistry (ABC) , 2012, DOI: 10.4236/abc.2012.22011
Abstract: Rhodopsin oligomerization and dissociation in vivo and under experimental conditions is an important topic both for a basic understanding of photoreceptor structure-function but also as a potential eye disease mechanism. In this study, to estimate a state rhodopsin after solubilization with mild and harsh detergents, we applied the native (blue native-PAGE, BN- PAGE) and denaturing electrophoresis (blue-urea- PAGE, BU-PAGE; blue-SDS-PAGE, BSDS-PAGE and SDS- PAGE). After blue BN-PAGE and BSDS- PAGE, rhodopsin and opsin, respectively, were presented in gels a major band of dimer with slight contents of higher oligomers without any traces of monomer, thus testifying in favor dimer-heteromeric state of frog rhodopsin in the photoreceptor membrane. Despite all oligomer bands gave positive staining with the rhodopsin-specific monoclonal antibodies (mAb), subsequent SDS-PAGE in combination with electroelution in denaturing conditions showed that stained bands are not homogenous and besides of opsin oli-gomers contain a small admixture of proteins with unknown function. Unfolding of opsin oligomers by solubilization in SDS, as compared with folded opsin in digitonin, induces their transition to a more compact conformation. It was manifested in a more rapid migration of opsin oligomers toward to anode. Cooling of digitonin/SDS mixed extracts at 4?C for 24 hours led to a partial reverse transition of unfolded opsin dimer to initial folded conformation, thus de- monstrating the entropic nature of this transition. Opsin monomer can be observed in the gels only after harsh dissociation of oligomers under BU-PAGE or SDS-PAGE. The electro elution of the individual opsin oligomers with denaturing buffer followed by SDS-PAGE resulted in dissociation of dimer to monomers. However, unexpectedly, the trimer was dissociated to a prevailing dimer and a small portion of monomer. The products dissociation of both opsin tetramer and pentamer are difficult to determine precisely, but they are neither monomer nor dimer. Dissociation data show that the degree of opsin oli- gomerization by unknown reasons affects the pattern of dissociation of its aggregates. Obtained in this paper data indicate a need for further detailed study the obscure mechanisms of aggregation-dissociation of rhodopsin.
Troponin based studies in search of a biomarker for cardiac arrest  [PDF]
Pasha Ghazal, Kaneez Fatima Shad, Nikhat Sidduiqui
Health (Health) , 2010, DOI: 10.4236/health.2010.21012
Abstract: Cardiac arrest is shown to be a cause of a large number of deaths not only in Pakistan but around the globe. The prevalence of this dis-ease demands identification of its etiology. The science of proteomics can be used to identify cardiac specific proteins. The subsequent over expression or under expression of these pro-teins can be utilized as targets not only for therapeutical interventions but also for identi-fying molecular signatures for Cardiac diseases. In context of a number of studies which have shown that the specificity of serum biomarkers like troponin (cTnI and cTnT) are questionable as they may also appear in serum in pathologi-cal conditions other than cardiac dysfunction, the search of a specific marker for cardiac arrest becomes imperative. In this study protein pro-filing of cardiac arrest patients was performed after its quantification through Bradford assay. SDS-PAGE and 2 DE techniques were used as to characterize proteins. The samples of the pa-tients prior to characterizing of proteins were subjected to lipid and cardiac enzymes profiling. The results of these investigations have shown an increase in almost all of these parameters by many folds from that of normal values. In addi-tion to this the samples were found out to be positive for troponin T which strongly confirms the incidence of the cardiac arrest. The results of SDS-PAGE exhibited the induction of three proteins of 100 kDa, 97 kDa and of 66 kDa with 100 kDa as the most highly expressed protein. In addition to that SDS-PAGE gels have shown the down regulation of 45 kDa protein, again indi-cating the changes as a result of cardiac arrest. 2DE gel patterns of cardiac arrest samples demonstrated higher number of protein spots as compare to control in the alkaline range, which might suggest their role in cardiac dysfunction. Therefore it can be concluded that this study may pave the grounds for identification of such proteins which can serve not only as potential therapeutical targets but also as candidate markers for accurate diagnosis of the disease.
Humic and Oxalic Acid Stimulates Grain Yield and Induces Accumulation of Plastidial Carbohydrate Metabolism Enzymes in Wheat Grown under Sandy Soil Conditions  [PDF]
Bakry Ahmed Bakry, Hattem M. El-Shabrawi, Mohamed A. Ahmed, Mohamed Abou-El-Lail
Agricultural Sciences (AS) , 2015, DOI: 10.4236/as.2015.61016
Abstract: Humic and oxalic acids have the effects of promoting plant growth. We test whether they are able to positively impact wheat yield under newly reclaimed sandy soil, where water deficiency negatively influences yield. Foliar application of humic acid and oxalic acid on two wheat cultivars, Gemiza-9 and Sakha-93, leads to overall better performance of the plants and increases the yield significantly, irrespective of the cultivar genetic background. However, Gemiza-9 surpassed Sak- ha-93 in grain yield parameters. The highest values of grain and protein yields/ha were obtained in both cultivars, when the plants were sprayed with a combination of 17 mg/L humic acid and (300 mg/L) oxalic acid. Humic and oxalic acid showed accumulative yield-promoting effect. To understand the mechanism by which humic and oxalic acids promoted grain yield, we performed SDS-PAGE followed by MS-MS-LC analyses. We identified a unique humic acid-induced 52 KDa band in Gemiza-9. The band contained three major proteins, Ribulose bisphosphate carboxylase large chain, ADP-glucose synthase and NADP-dependent glyceraldehyde-3-phosphate dehydrogenase (GAPN). Thus humic acid increased the activity of plastid enzymes involved in photosynthesis, sucrose biosynthesis and starched accumulation to improve the overall performance of the plant.
Optima of Trypsin-Catalyzed Hydrolysis and Its Inhibition Determined by SDS-PAGE  [PDF]
Xueke Zhou, Tingting Wang, Anjun Wang, Renqiang Li
Advances in Enzyme Research (AER) , 2016, DOI: 10.4236/aer.2016.41001
Abstract: SDS-PAGE was applied to determine trypsin activity and inhibition. After the hydrolysis by trypsin to substrate bovine serum albulnin (BSA) under different temperatures and pH, the hydrolysis degree of BSA was conducted using SDS-PAGE. From the quantitative analysis to the electrophoresis bands of BSA and its hydrolysis products in SDS-PAGE pattern, the change of trypsin activity was determined, and then the optimum temperature at 40°C and the optimum pH at pH 8.5 - 8.7 for trypsin activity were obtained. All the target bonds in BSA molecule could be hydrolyzed at the same time by trypsin. The inhibition was due to the binding of inhibitor to trypsin, which made it impossible for trypsin to touch the substrate protein. SDS-PAGE was demonstrated to be also an effect method for assaying the characteristics of trypsin activity and its inhibition.
Comparison of Haemophilus parasuis reference strains and field isolates by using random amplified polymorphic DNA and protein profiles
Emilie S Zehr, Dennis V Lavrov, Louisa B Tabatabai
BMC Microbiology , 2012, DOI: 10.1186/1471-2180-12-108
Abstract: The DNA and WCP lysate profiles of 15 reference strains and 31 field isolates of H. parasuis were analyzed using the Dice and neighbor joining algorithms. The results revealed unique and reproducible DNA and protein profiles among the reference strains and field isolates studied. Simpson’s index of diversity showed significant discrimination between isolates when three 10mer primers were combined for the RAPD method and also when both the RAPD and WCP lysate typing methods were combined.The RAPD profiles seen among the reference strains and field isolates did not appear to change over time which may reflect a lack of DNA mutations in the genes of the samples. The recent field isolates had different WCP lysate profiles than the reference strains, possibly because the number of passages of the type strains may affect their protein expression.
Cuantificacion de huevo en fideos secos según metodo electroforético (SDS PAGUE)
López,LB; Cellerino,K; Binaghi,MJ; Giacomino,MS; Valencia,ME;
Diaeta , 2011,
Abstract: the objective of this work was to use the sds-page method for the quantification of egg in dry noodles made with flour and added egg. 6 model systems (ms) of noodles containing 0.0, 1.0, 2.5, 4.0, 6.0 and 8.0% of egg powder were analyzed. total proteins with a buffer containing sodium dodecyl sulfate (sds) and 2-mercaptoethanol were extracted and electrophoresis with sds polyacrylamide gel was carried out. the relationships of the peak areas of the densitograms (egg and wheat) that allow a proper quantification of the percentage of egg added were established. 11 commercial samples in which quantification of egg was carried out were analyzed, with values ranging between <1.0% and 8.8% of egg powder. in the analyzed ms and in the commercial samples the cholesterol content (enzymatic method) was also determined and the egg content was determined by the method elisa. cholesterol values in the ms increased with the increase of the added egg powder, while in the samples, low levels of cholesterol were found for the lowest egg levels and high values for the highest egg levels. with the elisa method a correct quantification of egg up to 4.0% was achieved, but low results were obtained in the higher systems (6.0 and 8.0%). this could be due to the significant dilution that must be made from the extracts of the samples to determine their content. with respect to the commercial samples, in 8 similar values to those of electrophoresis were obtained, while in other 3, the values were different. the electrophoretic method is a useful tool for the detection and quantification of egg in these samples when you have ms with known powdered egg concentration.
Intraspecific variation in protein pattern of red scorpion (Mesobuthus tamulus, coconsis, pocock) venoms from Western and Southern India
Badhe, R. V.;Thomas, A. B.;Harer, S. L.;Deshpande, A. D.;Salvi, N.;Waghmare, A.;
Journal of Venomous Animals and Toxins including Tropical Diseases , 2006, DOI: 10.1590/S1678-91992006000400008
Abstract: red scorpions mesobuthus tamulus (coconsis, pocock) were obtained from different regions of west and south india (ratnagiri, chiplun and ahmednagar from maharashtra and chennai from tamil nadu, respectively). their venoms composition was analyzed using gel electrophoresis (sds-page). all venom samples shared six bands of 170, 80, 60, 57, 43, and 38 kda molecular weights. bands of 115 kda and 51.5 kda were characteristic of venoms obtained from red scorpions of chiplun region, and the 26kda band was absent in scorpion venom from tamil nadu. the separated protein band patterns suggest that the venoms from ratnagiri, ahmednagar and tamil nadu had high similarities in their biochemical composition but differed from that of chiplun region. these data were also supported by the jaccard (j) index. the j value was 0.33 for venom obtained from ratnagiri-ahmednagar, 0.31 for venom from ratnagiri-tamil nadu, and 0.3 for venom from ratnagiri-chiplun region. this suggests the existence of genetic variation among the different strains of red scorpion in western and southern india. the antiserum produced by haffkine biopharmaceuticals corporation ltd. completely neutralized proteins of venoms from all the regions studied.
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