Displaying publications 1 - 20 of 197 in total

  1. Amid M, Abd Manap MY
    Food Chem, 2014 Dec 15;165:412-8.
    PMID: 25038694 DOI: 10.1016/j.foodchem.2014.03.133
    An amylase enzyme from pitaya peel was purified 234.2-folds with 72.1% recovery using ammonium sulphate precipitation, gel filtration and ion exchange chromatography. Gel filtration chromatography and SDS-PAGE revealed that the enzyme is monomeric with a molecular weight of 42.1kDa. The apparent Km and Vmax of the amylase were 2.7 mg/ml and 34.30 u/min/mg of protein, respectively. The enzyme was highly active and stable over a wide pH range from pH 3 to pH 11.0, with optimum activity being observed at pH 5.0. The enzyme was highly selective for soluble starch, amylopectin, glycogen and pulullan. The purified amylase did not require calcium and displayed extreme stability with regard to surfactants and oxidising agents. EDTA, a powerful chelating agent, did not have any significant effect on the stability of the enzyme. Such characteristics have not been previously reported for this type of enzyme from fruit peel. This enzyme, which possesses unique properties, could be widely used in different types of industries, especially in food and biotechnological applications.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/methods*
  2. Chew FN, Tan WS, Ling TC, Tan CS, Tey BT
    Anal. Biochem., 2009 Jan 15;384(2):353-5.
    PMID: 18952038 DOI: 10.1016/j.ab.2008.10.010
    Green fluorescent protein (GFP) is a versatile reporter protein and has been widely used in biological research. However, its quantitation requires expensive equipment such as a spectrofluorometer. In the current study, a gel documentation imaging system using a native polyacrylamide gel for the quantitation of GFP was developed. The assay was evaluated for its precision, linearity, reproducibility, and sensitivity in the presence of Escherichia coli cells and was compared with the spectrofluorometric method. Using this newly established, gel-based imaging technique; the amount of GFP can be quantified accurately.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/methods*
  3. Gam LH, Latiff A
    Int. J. Biol. Sci., 2005;1(3):103-9.
    PMID: 16094462
    The microheterogeneity property of hCG with regards to its sialic acid contents resulted in variable mobility of the glycoprotein in SDS-PAGE. The intact hCG molecule is composed of two dissimilar subunits, namely alpha- and beta-subunits. The identification of hCG bands in SDS-PAGE was accomplished by the immunoblotting experiment, whereby the antibody directed toward the specific region of beta-subunit of hCG was used. The data shows that the different mobility of intact hCG was attributed to the different degree of desialylation of the glycoprotein. Nevertheless, unlike the intact hCG, the mobility of its beta-subunit was not affected by its variety sialic acid content. This characteristic of beta-hCG is beneficial when semi-quantification of total hCG is required. Quantification of hCG using the HPLC-reversed phase C18 analytical column is not possible as the glycoprotein was eluted in multiple fractions at different retention times. The identification of denatured hCG (HPLC eluted fractions) was carried out by immunoblotting experiment whilst immunoassay technique failed to detect its presence in any fraction.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/methods*
  4. Nur Azira T, Che Man YB, Raja Mohd Hafidz RN, Aina MA, Amin I
    Food Chem, 2014 May 15;151:286-92.
    PMID: 24423534 DOI: 10.1016/j.foodchem.2013.11.066
    The study was aimed to differentiate between porcine and bovine gelatines in adulterated samples by utilising sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) combined with principal component analysis (PCA). The distinct polypeptide patterns of 6 porcine type A and 6 bovine type B gelatines at molecular weight ranged from 50 to 220 kDa were studied. Experimental samples of raw gelatine were prepared by adding porcine gelatine in a proportion ranging from 5% to 50% (v/v) to bovine gelatine and vice versa. The method used was able to detect 5% porcine gelatine added to the bovine gelatine. There were no differences in the electrophoretic profiles of the jelly samples when the proteins were extracted with an acetone precipitation method. The simple approach employing SDS-PAGE and PCA reported in this paper may provide a useful tool for food authenticity issues concerning gelatine.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/methods*
  5. Normah, Ismail, Ezzana Zuraini, Zainuddin
    Scientific Research Journal, 2015;12(1):1-12.
    Proteases were extracted from starfruit at maturity Index 2 (unripe, light green) and Index 7 (very ripe, orange) and partially purified using acetone and 40% ammonium sulfate precipitations. Higher yield and proteolytic activity were observed for proteases purified using acetone than 40% ammonium sulfate. As for maturity index, yield and protein concentration of proteases from Index 2 were higher than those from Index 7. SDS-PAGE result showed intense bands for acetone proteases while a distinct band at 50 kDa was observed in all the proteases. Enzyme activity decreased during the seven days storage at 4°C with minimum relative activity of 70% achieved for acetone proteases at day seven. This study suggested that acetone precipitation is more effective method for purifying starfruit protease based on the yield and proteolytic activity compared to using 40% ammonium sulphate precipitation. In order to obtain higher protein concentration and proteolytic activity, starfruit at the unripe stage, Index 2 is a better raw material than Index 7 to be used for protease production.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  6. Chew FN, Tan WS, Tey BT
    J. Biosci. Bioeng., 2011 Feb;111(2):246-8.
    PMID: 21036662 DOI: 10.1016/j.jbiosc.2010.10.004
    A gel imaging method was employed to quantitate the GFP that had been subjected to denaturation and degradation treatments. This method is able to differentiate the nativity of GFP by relating the observed changes in the position of fluorescent bands which is unable to be detected using the spectrofluorometric method.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  7. Onsa GH, bin Saari N, Selamat J, Bakar J
    J. Agric. Food Chem., 2000 Oct;48(10):5041-5.
    PMID: 11052775
    Latent polyphenol oxidase (LPPO), an enzyme responsible for the browning reaction of sago starches during processing and storage, was investigated. The enzyme was effectively extracted and partially purified from the pith using combinations of nonionic detergents. With Triton X-114 and a temperature-induced phase partitioning method, the enzyme showed a recovery of 70% and purification of 4. 1-fold. Native PAGE analysis of the partially purified LPPO revealed three activity bands when stained with catechol and two bands with pyrogallol. The molecular masses of the enzymes were estimated by SDS-PAGE to be 37, 45, and 53 kDa. The enzyme showed optimum pH values of 4.5 with 4-methylcatechol as a substrate and 7.5 with pyrogallol. The LPPO was highly reactive toward diphenols and triphenols. The activity of the enzyme was greatly enhanced in the presence of trypsin, SDS, ethanol, and linoleic acid.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  8. Hoe, S.Z., Pendek, R., Lam, S.K., Rahim, Z.H.A.
    Ann Dent, 1997;4(1):-.
    Human saliva contains a large number of proteins which can be separated using polyacrylamide gel electrophoresis (PAGE). In this study the protein profiles of whole saliva of diabetic and non-diabetic were compared. Considerable variations between individuals in the protein profiles were observed. The saliva from diabetic patients appeared to have more of proline-rich protein bands in the molecular weight region below 56 KOa. Further investigations using individual gland saliva should be carried out.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  9. Mohd Rosni, S., Fisal, A., Azwan, A., Chye, F. Y., Matanjun, P.
    It is crucial to determine several protein-related parameters at the initial stages of proteomic analysis of any biological samples. In this study, crude protein content, total soluble protein, total phenolic content and the SDS-PAGE profile of fifteen varieties of seaweed from Semporna, Sabah, Malaysia were analysed. The crude protein, total soluble protein and total phenolic content of all seaweed samples were in the range of 3.99 to 13.18 % of dry weight, 0.52 to 1.45 mg/mL in acetone dried powder samples and 8.59 to 48.98 mg PGE/g dry weight, respectively. In general, the differences (crude protein, total soluble protein and total phenolic content) among all fifteen varieties of seaweeds were significant (p< 0.05). There was also a strong positive correlation between crude protein and total soluble protein concentration (Pearson’s Correlation Coefficient (r)=0.923; p=0.01) in these fifteen varieties of seaweed. A distinctive protein pattern was observed in the SDS-PAGE gels between three different seaweed classes of green, red and brown colours. All of these results are important in sample preparations (extractions) before furthering proteomic analysis in order to identify and characterize seaweed proteomes.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  10. Jamilah, B., Umi Hartina, M.R., Mat Hashim, D., Sazili, A.Q.
    The properties of collagens from Barramundi (Lates calcarifer) skin obtained by acid solubilized (control), pepsin and papain aided extractions were investigated. The yields of collagens (dry weight basis) for acid solubilized, pepsin and papain aided extractions were 8.1, 43.6 and 44.0%, respectively. The collagens were generally colorless although collagens from the enzymes aided-extractions were slightly darker. Based on the e-nose evaluation, the collagens were considered odorless. The pH of all the collagens was in the vicinity of 3; however, those extracted with papain had significantly higher pH. The polypeptide profiles obtained in the SDS-PAGE analysis for pepsin extracted collagen were similar to those of acid solubilized collagens. Papain extracted collagen had distinctly different SDS-PAGE pattern. All the extracted collagens were of type 1 with apparent peptides molecular weight distribution of 37 to 250 kDalton. They had high solubility in pH 2 to 5 and increasing NaCl concentration up to 6%.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  11. Mohd Khalizan Sabullah, Azlan Jualang Gansau, Mohd Rosni Sulaiman, Fisal Ahmad
    Observations on the effects of copper on the liver proteome of Puntius javanicus based on the
    one dimensional PAGE was carried out. The liver was dissected from each fish, which was
    separately treated with different concentrations of copper sulfate ranging from 0.1 to 5.0 mg/L.
    The livers were extracted and one dimensional PAGE was performed under nonreducing
    (native) and reducing (SDS)-PAGE. Several bands were resolved in the native PAGE with
    probable candidates for the effect of copper observed showing an increased in the expression
    and downregulation strongly associated with increasing copper concentrations. This study
    showed that high concentrations of copper significantly alters P. javanicus liver at the proteome
    level, and preliminary screening based on one dimensional PAGE is considered rapid and
    simple to assess the toxicity effect of copper before more advanced and extensive assesment
    with a second dimensional PAGE is carried out.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  12. Mohd Fakharul Zaman Raja Yahya, Hasidah Mohd Sidek
    Kajian ini melibatkan pemantauan perkembangan parasitemia dan taburan morfologi Plasmodium berghei sewaktu infeksi parasit dalam mencit, serta penentuan kesan infeksi P. berghei ke atas pengisyaratan MAP kinase eritrosit perumah. Analisis mikroskop ke atas slaid calitan darah terwarna-Giemsa yang disediakan daripada mencit terinfeksi-P. berghei (strain PZZ1/00) menunjukkan darjah parasitemia mencapai sehingga 70% dalam masa dua minggu selepas penyuntikan parasit. Morfologi cecincin dan trofozoit parasit dicerap dengan jelas sepanjang tempoh infeksi manakala morfologi skizon parasit hanya dicerap dengan ketara selepas hari ketiga selepas penyuntikan parasit. Pemblotan Western [antibodi primer: anti-MAP kinase (ERK-1/2 tak terfosfat) monoklon; antibodi sekunder: anti-IgG, poliklon terkonjugat-HRP] ke atas protein sitosol eritrosit terinfeksi-P. berghei (70% parasitemia) susulan pemisahan SDS-PAGE menunjukkan bahawa keamatan protein imunoreaktif-MAP kinase eritrosit berberat molekul 42 dan 44 kDa didapati meningkat secara signifikan (p<0.05) pada 70% iaitu peningkatan sebanyak 21.5% dan 22.3% masing-masing berbanding sampel kawalan tanpa infeksi. Samada kesan infeksi P. berghei (70% parasitemia) ke atas pengisyaratan MAP kinase perumah ini berkaitan dengan pengaktifan enzim ini perlu dikaji dengan lebih lanjut.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  13. Rasli, H.I., Sarbon, N.M.
    Enzymatic hydrolysis of proteins is an important bioprocess method to prepare bioactive peptides with many functionality and health benefits. The aims of the present work were to prepare and determine the physicochemical characteristics of gelatine hydrolysate from skin of shortfin scad (SSGH) via hydrolysis using alcalase. Analyses on chemical composition, molecular weight by SDS PAGE, protein concentration, amino acid composition, Fourier Transform Infrared Spectroscopic features, and solubility of SSGH were thus performed. The yield of SSGH obtained was 51.01% (d.b.). The chemical compositions of SSGH for moisture, protein, fat, and ash were 13.82%, 90.05%, 1.95%, and 12.48%, respectively. SSGH showed low molecular weight (
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  14. Chew FN, Tan WS, Ling TC, Tey BT
    Electrophoresis, 2009 Sep;30(17):3017-23.
    PMID: 19685471 DOI: 10.1002/elps.200900246
    Mechanical and non-mechanical breakages of bacterial cells are usually the preliminary steps in intracellular protein purification. In this study, the recombinant green fluorescent protein (GFP) was purified from intact Escherichia coli cells using preparative PAGE. In this purification process, cells disruption step is not needed. The cellular content of E. coli was drifted out electrically from cells and the negatively charged GFP was further electroeluted from polyacrylamide gel column. SEM investigation of the electrophoresed cells revealed substantial structural damage at the cellular level. This integrated purification technique has successfully recovered the intracellular GFP with a yield of 82% and purity of 95%.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/methods*
  15. Lee CY, Hemingway J, Yap HH, Chong NL
    Med. Vet. Entomol., 2000 Mar;14(1):11-8.
    PMID: 10759307
    The possible insecticide resistance mechanisms of four Malaysian field-collected strains of the German cockroach, Blattella germanica (Linnaeus) (Dictyoptera: Blattellidae), were characterized with biochemical assays and native polyacrylamide gel electrophoresis (PAGE). Elevated esterase activity (at low to moderate frequency) and altered acetylcholinesterase (low frequency) were detected in all field strains, while elevated glutathione S-transferase levels were present in only two strains. Seven esterase bands were separated by native PAGE; a greater intensity occurred in three bands in the resistant strains compared to the susceptible strain. Inhibition studies using specific inhibitors on polyacrylamide gels suggested that the slowest of these three esterases is a cholinesterase, while the other two are carboxylesterases with a preference for beta- over alpha-naphthyl acetate.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/veterinary
  16. Lee HX, Ahmad F, Saad B, Ismail MN
    Prep. Biochem. Biotechnol., 2017 Nov 26;47(10):998-1007.
    PMID: 28857669 DOI: 10.1080/10826068.2017.1365250
    Date fruits are well known to be very nutritious. Nevertheless, the protein contents of the fruit, particularly the seed and flesh, are still understudied, largely due to their difficult physical characteristics. This study was conducted to compare three different protein extraction methods which were the trichloroacetic acid (TCA)-acetone (TCA-A), phenol (Phe), and TCA-acetone-phenol (TCA-A-Phe), and to perform proteomic analysis on date palm seed and flesh. Phe extraction method showed the highest protein yields for both seed (8.26 mg/g) and flesh (1.57 mg/g). Through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Phe, and TCA-A-Phe extraction methods were shown to be efficient in removing interfering compounds and gave well-resolved bands over a wide range of molecular weights. Following liquid chromatography-tandem mass spectrometry analysis, about 50-64% of extracted proteins were identified with known functions including those involved in glycolysis, Krebs cycle, defense, and storage. Phe protein extraction method was proven to be the optimal method for date flesh and seed.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/methods
  17. Tan KY, Liew ST, Tan QY, Abdul-Rahman FN, Azmi NI, Sim SM, et al.
    Toxicon, 2019 Mar 15;160:55-58.
    PMID: 30797900 DOI: 10.1016/j.toxicon.2019.02.010
    Gel filtration chromatography and gel electrophoresis revealed minimal protein degradation in lyophilized antivenoms which were 2-year expired (Hemato Polyvalent, Neuro Polyvalent; Thailand) and 18-year expired (Hemato Bivalent, Neuro Bivalent; Taiwan). All expired antivenoms retained immunological binding activity, and were able to neutralize the hemotoxic or neurotoxic as well as lethal effects of the homologous snake venoms. The findings show that antivenoms under proper storage conditions may remain relatively stable beyond the indicated shelf life.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel/methods
  18. Chua LS, Abdul-Rahman N, Rosidi B, Lee CT
    Nat. Prod. Res., 2013 Mar;27(4-5):314-8.
    PMID: 22468741 DOI: 10.1080/14786419.2012.676552
    A water extraction method has been used to extract plant proteins from the roots of Eurycoma longifolia harvested from Perak and Pahang, Malaysia. On the basis of the spectroscopic Bradford assay, Tongkat Ali Perak and Pahang contained 0.3868 and 0.9573 mg mL(-1) of crude protein, respectively. The crude proteins were separated by one dimensional 15% sodium dodecyl sulphate polyacrylamide gel electrophoresis into two (49.8 and 5.5 kD) and four (49.8, 24.7, 21.1 and 5.5 kD) protein spots for Tongkat Ali Perak and Pahang, respectively. Isoleucine was present in the highest concentration significantly. Both plant samples showed differences in the mineral and trace element profiles, but the minerals calcium, magnesium and potassium were present in the highest concentration. The highly concerned toxic metals such as arsenic and lead were not detected.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  19. Tan CY, Rahman RN, Kadir HA, Tayyab S
    Acta Biochim. Pol., 2011;58(3):405-12.
    PMID: 21887412
    Bacillus licheniformis α-amylase (BLA) was chemically modified using 100-fold molar excess of succinic anhydride over protein or 0.66 M potassium cyanate to obtain 42 % succinylated and 81 % carbamylated BLAs. Size and charge homogeneity of modified preparations was established by Sephacryl S-200 HR gel chromatography and polyacrylamide gel electrophoresis. Conformational alteration in these preparations was evident by the larger Stokes radii (3.40 nm for carbamylated and 3.34 nm for succinylated BLAs) compared to 2.43 nm obtained for native BLA. Urea denaturation results using mean residue ellipticity (MRE) as a probe also showed conformational destabilization based on the early start of transition as well as ΔG(D)(H(2)O) values obtained for both modified derivatives and Ca-depleted BLA. Decrease in ΔG(D)(H(2)O) value from 5,930 cal/mol (for native BLA) to 3,957 cal/mol (for succinylated BLA), 3,336 cal/mol (for carbamylated BLA) and 3,430 cal/mol for Ca-depleted BLA suggested reduced conformational stability upon modification of amino groups of BLA or depletion of calcium. Since both succinylation and carbamylation reactions abolish the positive charge on amino groups (both α- and ε- amino), the decrease in conformational stability can be ascribed to the disruption of salt bridges present in the protein which might have released the intrinsic calcium from its binding site.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
  20. Kuan YH, Bhat R, Karim AA
    J. Agric. Food Chem., 2011 Apr 27;59(8):4111-8.
    PMID: 21401213 DOI: 10.1021/jf104050k
    The physicochemical and functional properties of ultraviolet (UV)-treated egg white protein (EW) and sodium caseinate (SC) were investigated. UV irradiation of the proteins was carried out for 30, 60, 90, and 120 min. However, the SC samples were subjected to extended UV irradiation for 4 and 6 h as no difference was found on the initial UV exposure time. Formol titration, SDS-PAGE, and FTIR analyses indicated that UV irradiation could induce cross-linking on proteins and led to improved emulsifying and foaming properties (P < 0.05). These results indicated that the UV-irradiated EW and SC could be used as novel emulsifier and foaming agents in broad food systems for stabilizing and foaming purposes.
    Matched MeSH terms: Electrophoresis, Polyacrylamide Gel
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