• Title/Summary/Keyword: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Electrophoretic analysis of the major proteins of race horse erythrocyte membrane: Their relation to fast erythrocyte sedimentation rate (경주 적혈구막 단백의 전기영동법에 의한 분석 -빠른 적혈구 침강 속도와의 관계-)

  • Bahk, Yeong-woo;Lee, Si-yeong
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.259-264
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    • 1991
  • The proteins of the race horse erythrocyte membrane were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate(SDS-PAGE), and their relations to the fast erythrocyte sedimentation rate(ESR) of the race horse were investigated. The erythrocyte sedimentation rate of race horse were very fast compared with the human one(33 times <$90^{\circ}-plastic-ESR/30m$> and 25 times <$90^{\circ}-micro-ESR/30m$> as fast as the human one) are reported previously. Although the general protein profiles of the race horse erythrocyte membranes were almost similar to that of human, band 3 content was showing higher in race horse (34.7%) than in human (25.3%). The glycoprotein profiles of the race horse erythrocyte membranes revealed by periodic acid Schiff's(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycoprotein) present in human erythrocyte memo brane were almost absent from the Holstein and race horse erythrocyte membranes, but PAS-2 was more in only race horse from that of human. Instead, the bovine erythrocyte membranes showed a strong PAS-B near the origin of the electrophorograms and the race horse erythrocyte membranes showed a strong PAS-negative band near the end of the electrophorograms, which is named as PAS-E in this study. These results suggest that the fast sedimentation rate of race horse erythrocyte is due in part to the presence of more band 3 protein fraction and PAS-E glycoproteins in the race horse erythrocytes.

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Low Protein Digestibility of Beef Puree in Infant In Vitro Digestion Model

  • Lee, Seonmin;Jo, Kyung;Hur, Sun Jin;Choi, Yun-Sang;Kim, Hyun-Joo;Jung, Samooel
    • Food Science of Animal Resources
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    • v.39 no.6
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    • pp.1000-1007
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    • 2019
  • This study investigated protein digestibility of beef puree in infant and adult in vitro digestion models. The simulated digestive juices for infant and adult were prepared. Protein digestibility of beef puree was calculated in the gastric and gastrointestinal compartments. The 10% trichloroacetic acid soluble nitrogen and α-amino group contents of gastric digesta were lower in the infant in vitro digestion model than those in the adult in vitro digestion model (p<0.05). In addition, the gastrointestinal digesta from the infant in vitro digestion model had lower value of the 10% trichloroacetic acid soluble nitrogen and α-amino group contents than those of the adult in vitro digestion model (p<0.05). The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the remarkable bands of actin and myosin light chain B were found in the digesta of beef puree from the infant in vitro digestion model. The results of this study revealed the lower protein digestibility of beef puree in infants compared to that in adults. Therefore, the development of ways to increase digestibility of meat protein can improve the nutritional quality of meat products for infants.

Postmorten Changes in the Myofibrillar Proteins of Alkaline and Acid Rigor Rabbit Muscle during Storage (사후강직형태에 따른 근원섬유 단백질의 이화학적 성질변화)

  • Kim, Dae-Gon;Sung, Sam-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.6
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    • pp.712-718
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    • 1992
  • This experiment was carried out to investigate some physico-chemical properties of myofibrillar proteins isolated from alkaline and acid rigor muscles of rabbit. The degree of fragmentation was about 0.35 at 3 days of postmortem in acid rigor muscle fiber, whereas it was only 0.3 at 7 days of postmortem in alkaline rigormuscle fiber. $Mg^{2+}$-activated ATPase activities of actomyosin were increased to the highest level at 1st day in acid rigor and at 3rd day in alkaline rigor muscle fiber. $Ca^{2+}$-activated ATPase activities of actomyosin were slightly increased at postmortem. $Mg^{2+}$-, and $Ca^{2+}$-activated ATPase activities were higher in alkaline rigor muscle than those in acid rigor muscle at postmortem. Solubility of actomyosin increased with postmortem but no differences were observed in rigor types. SDS-PAGE(sodium dodecyl sulfate polyacrylamide gel electrophoresis) band patterns of myofibrils did not show significant differences between rigor types, but newly 30.000 dalton proteins were appeared at 3 day postmortem in both rigor types.

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Neutralization of Human Papillomavirus by Specific Nanobodies Against Major Capsid Protein L1

  • Minaeian, Sara;Rahbarizadeh, Fatemeh;Zarkesh-Esfahani, Sayyed Hamid;Ahmadvand, Davoud;Broom, Oliver Jay
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.721-728
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    • 2012
  • The human papillomavirus (HPV) is the main cause of cervical cancer in developing countries. Rapid diagnosis and initiation of treatment of the HPV infection are critical. Various methods have been employed to reduce the immunogenicity of antibodies targeting HPV serotypes. Nanobodies are the smallest fragments of naturally occurring single-domain antibodies with their antigen-binding site compromised into a single domain. Nanobodies have remarkable properties such as high stability, solubility, and high homology to the human VH3 domain. In this study, a phagemid library was employed to enrich for nanobodies against the L1 protein of the human papilloma virus. Binding reactivity of the selected clones was evaluated using phage enzyme-linked immunosorbent assay (phage-ELISA). Finally, two nanobodies (sm5 and sm8) with the best reactivity against the Gardasil vaccine and the purified HPV-16 L1 protein were expressed and purified using a $Ni^+$-NTA column. The accuracy of expression and purification of the nanobodies was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting assays. In vitro studies demonstrated that neutralization was achieved by the selected nanobodies. The ease of generation and unique features of these molecules make nanobodies promising molecules for the new generation of HPV diagnosis and therapy.

Purification and Characterization of Thermostable Agarase from Bacillus sp. BI-3, a Thermophilic Bacterium Isolated from Hot Spring

  • Li, Jiang;Sha, Yujie;Seswita-Zilda, Dewi;Hu, Qiushi;He, Peiqing
    • Journal of Microbiology and Biotechnology
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    • v.24 no.1
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    • pp.19-25
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    • 2014
  • An extracellular agarase was purified from Bacillus sp. BI-3, a thermophilic agar-degrading bacterium isolated from a hot spring in Indonesia. The purified agarase revealed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular mass of 58 kDa. The optimum pH and temperature of the agarase were 6.4 and $70^{\circ}C$, respectively. The activity of the agarase was stable at high temperatures, and more than 50% activity was retained at $80^{\circ}C$ for 15 min. Furthermore, the enzyme was stable in the pH range of 5.8-8.0, and more than 60% of the residual activity was retained. Significant activation of the agarase was observed in the presence of $K^+$, $Na^+$, $Ca^{2+}$, $Mg^{2+}$, and $Sr^{2+}$; on the other hand, $Ba^{2+}$, $Zn^{2+}$, $Cu^{2+}$, $Mn^{2+}$, $Co^{2+}$, $Fe^{2+}$, and EDTA inhibited or inactivated the enzyme activity. The components of the hydrolytic product analyzed by thin-layer chromatography showed that the agarase mainly produced neoagarobiose. This study is the first to present evidence of agarolytic activity in aerobic thermophilic bacteria.

Comparison of IgE induction in mice by pollens from three pine tree species

  • Kim, Seo-Yoong;Oh, In-Bo;Choi, Kee-Ryong
    • Journal of Ecology and Environment
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    • v.41 no.9
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    • pp.265-270
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    • 2017
  • Background: Over the years, pine pollens have been excluded as an allergen due to its relatively large size, low protein content, and waxy hydrophobic layer, despite their abundance. However, recent studies suggest the possibilities of pine pollens being allergens, and it has been reported that allergy symptoms were highly prevalent in areas with considerably large pine forests and high possibility of exposure to the pollen. Therefore, we conducted a comparative analysis of the allergenicities of the pollens from the dominant species of Korean pines, red pine (Pinus densiflora), black pine (Pinus thunbergii), and pitch pine (Pinus rigida), in mice. Methods: The protein composition of the pollens from the three pine species was compared via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The pine pollens and proteins extracted from the pollens were introduced to BALB/c mice by nasal inhalation and application to exposed skin and the IgE produced by the mice were extracted from blood and analyzed via ELISA. Results: SDS-PAGE showed differing protein compositions of the pollens of the three pine species. Analysis of blood IgE compositions showed a similar amount of IgE produced when pollens were applied to skin. In contrast, when mice inhaled the pollens, P. densiflora was shown to induce significantly more IgE production than those of the other two species. Conclusions: The experimental results demonstrate that the pollens of all three South Korean pine species induce IgE production, and this production was more pronounced when the pollens were inhaled than when they were applied to the skin. Of the three species, the pollen of P. densiflora was found to induce the highest level of IgE production.

Changes of Corn Proteins and Lipids induced by Thermal Processing (옥수수 가열가공처리에 의한 단백질 및 지질성분의 변화)

  • Cho, Sung-Hwan;Yoon, Zoo-Lk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.18 no.3
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    • pp.287-299
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    • 1989
  • This research was conducted in order to investigate thermal stability and nutritional value of corn lipids and proteins during thermal processing. The lipids of raw and popped corn were fractionated and analyzed by column and gas chromatography. The effect of thermal processing on corn proteins was examined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and amino acid analysis. There was no remarkable change in proximate compositions during thermal processing. The lipid fractions obtained by silicic acid column chromatography were composed of neutral lipid(93.5%), glycolipid(3.8%), and phospholipid(2.7%), Although the thermal processing showed the increase in the ratio of unsaturated/saturated fatty acid, there was no significant differences in the lipid composition between raw and popped corn. Most of each protein fractions had lower molecular weight than 25,000 dalton and albumin fractions were distributed in the molecular weight range 11,500-94,000 daltons. Popped corn proteins did not show marked differences in their electrophoretic migrations when compared with raw corn proteins.

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Production of Recombinant Protein, Human Stem Cell Factor, Using Insect Cell Line

  • Park, Sang-Mi;Kwon, Ki-Sang;Goo, Tae-Won;Yun, Eun-Young;Kang, Seok-Woo;Kim, Sung-Wan;Yu, Kweon;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.37-45
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    • 2009
  • Insect cell cultures have become important tools in the production of biological substances for use in a variety of research, human and veterinary medicine, and pest control applications. These applications often require the introduction of foreign DNA into the cells and have generally used methods originally developed for use with human and other mammalian cell cultures. While these methods can be successfully employed, they are often less efficient with insect cells and frequently involve complex procedures or require specialized equipment. Even when they do work, they may require substantial modification because of differences in the culture medium or growth patterns of insect cells. In this study, We have optimized transfection conditions of Sf9 cell line using insect expression vector pIZT/V5-His which expresses green fluorescent protein effectively. Human stem cell factor (hSCF) is a glycoprotein that plays a key role in hematopoiesis acting both as a positive and negative regulator, often in synergy with other cytokines. It also plays a key role in mast cell development, gametogenesis, and melanogenesis. It can exist in membrane-bound form and in proteolytically released soluble form. As determined by an enzyme-linked immunosorbent assay performed, hSCF level in supernatant averaged 995ng/ml. The human hSCF was partially purified by immunoaffinity chromatography and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The results show that the hSCF has N-linked carbohydrate and corresponds to the soluble form, at or about 223 amino acids in length. The findings suggest functional importance for soluble hSCF in cells.

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The Effect of Salivary Gland of Streptozotocin Induced Diabetic Rats by Stress (스트레스가 Streptozotocin유도 당뇨백서의 타액선에 미치는 영향)

  • 김홍모;전양현;홍정표
    • Journal of Oral Medicine and Pain
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    • v.22 no.1
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    • pp.65-80
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    • 1997
  • Cytochrome P450 is an oxidase involved in oxidation of alcohol and is known to be an activator of carcinogen. The present study was perfomed to analyze the effect of diabetes and cold stress on the expression of Cytochrome P450 IIE1(CYPIIE1) in the liver and salivary glands in rats by an immunoblot analysis. Sixty three divided into 4 groups; 1) 20 rats belonging to group I were allowed diabetes (40mg/kg. I.V.) 2) 20 rats of group II were bathed in cold water for 30 seconds twice a day 3) 20 rats comprising group III were received diabetes and cold stress as described above 4) 3 rats of group IV were selected as a control. The rats were sacrificed at the end of the same day 1, 2, 3, 4 weeks experiment. The liver and parotid glands were removed and stored at $-70^{\circ}C$ until use. The stored organs were homogenized for 10 seconds and the supernatants were obtained by centrifugation. The proteins of the supernatants were separated by sodium dodecyl sulfate- polyacrylamide gel electrophoresis and subjected to Western blotting. The blotted membranes were incubated with polyclonal antibodies to CYPIIE1. And sepimens were observed with light microscope also under the Hematoxillin-Eosin staining. The obtained results were as follows : 1. In diabetes group, acini had changed to degeneration severely 1 week experiment, but repaired gradually in lapse of time. 2. In diabetes group, septal connective tissue had changed to degeneration little by little from 1 week after experiment, and progressed severely in lapse of time. 3. In stress group, acini had not changed remarkably, but slightly separated each other 3 weeks after experiment. 4. In diabetes and stress group, histological feature had changed remarkably campare with in the group of diabetes only. 5. In all experimental group, CTPIIE1 had expressed remarkably in the liver tissue, but not in the parotid gland tissues. 6. In diabetes and stress group, CYPIIE1 had expressed remarkably campare with in the group of diabetes only.

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Genetic variation of 7S and 11S globulins in soybean seed (콩 종실 단백질의 유전변이)

    • Korean Journal of Plant Resources
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    • v.12 no.3
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    • pp.198-203
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    • 1999
  • 7S and 11S globulins are two major storage proteins in soybean seed. For improving the quality of soybean seed protein, an increase of 11S/7S ratio would be a desirable objective because 11S globulin contains much more sulfur-containing amino acids than 7S globulin. In this study, six soybean varieties grown at three locations were used for genetic variation analysis of 7S and 11S globulins. It was possible to screen the soybean genotypes having aberrant subunit compositions of the two globulins by a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). So, heritabilities, genotypic and phenotypic correlations among eight globulin fraction contents of soybean seeds were estimated. The mean value of 7S and 11S globulin fraction contents were 38.9% and 61.1%, respectively, and the ratio of 7S to 11S globulin ranged from 0.58 to 0.74. The high heritability value was found in $\beta$ subunits but the values of acidic and basic subunits were relatively low. Genotypic correlations were higher than the corresponding phenotypic correlations in most of globulin subunit contents. $\beta$ subunits was negatively correlated with $\alpha$ and $\alpha$' subunits among 7S fractions, while no significant correlation between $\alpha$ and $\alpha$' subunits could be found In case of 11S fractions, acidic and basic subunits exhibited no genotypic but negative phenotypic correlation.

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