• Title/Summary/Keyword: Tris-buffer solution

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Interaction of ct-DNA with 2,4-Dihydroxy Salophen

  • Azani, Mohammad-Reza;Hassanpour, Azin;Bordbar, Abdol-Khalegh;Mirkhani, Valiollah
    • Bulletin of the Korean Chemical Society
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    • v.30 no.9
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    • pp.1973-1977
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    • 2009
  • In the present study, at first, 2,4-Dihydroxy Salophen (2,4-DHS), has been synthesized by combination of 1, 2-diaminobenzene and 2,4-dihydroxybenzaldehyde in a solvent system. This ligand containing meta-quinone functional groups were characterized using UV-Vis and IR spectroscopies. Subsequently, the interaction between native calf thymus deoxyribonucleic acid (ct-DNA) and 2,4-DHS, was investigated in 10 mM Tris/HCl buffer solution, pH 7.2, using UV-visible absorption and fluorescence spectroscopies, thermal denaturation technique and viscosity measurements. From spectrophotometric titration experiments, the binding constant of 2,4-DHS with ct-DNA was found to be (1.1 ${\pm}\;0.2)\;{\times}\;10^4\;M^{-1}.$ The fluorescence study represents the quenching effect of 2,4-DHS on bound ethidium bromide to DNA. The quenching process obeys linear Stern-Volmer equation in extended range of 2,4-DHS concentration. Thermal denaturation experiments represent the increasing of melting temperature of DNA (about 3.5 ${^{\circ}C}$) due to binding of 2,4-DHS. These results are consistent with a binding mode dominated by interactions with the groove of ct-DNA.

Determination of Horseradish Peroxidase (HRP) using an Enhanced Chemiluminescence Assay (증강 화학발광 기법을 이용한 horseradish peroxidase(HRP)의 검량)

  • Kim, Wongee;Kim, Keunhan;Lee, Seungmok
    • Journal of Korean Society on Water Environment
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    • v.25 no.1
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    • pp.84-89
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    • 2009
  • Our nation's water resources remain susceptible to contamination by phenolic agrichemicals. These compounds can be toxic to a variety of organisms including humans. Their disposal is restricted in many countries with strict limits for acceptable concentrations in drinking water. Enzyme-mediated in situ stabilization has been advocated as an approach for the treatment of phenolic compounds in soils and groundwater. This study reports the development of a new approach to quantify the activity of the HRP enzyme in aqueous systems. The method is based on the coupled processes of energy transfer and enhanced chemiluminescence using a luminol-$H_2O_2$-HRP system. In this study, the effects of solution pH, ionic strength and aqueous concentrations of HRP, $H_2O_2$ and enhancer were evaluated on the p-iodophenol-enhanced, HRP-catalyzed chemiluminescence reaction intensity in Tris-HCl buffer. All assay components were found to affect the maximum chemiluminescene intensity. The calibration curve for HRP showed the linear relationship with maximum light intensity.

Physical Properties and Hydroxyapatite Formation of Low Alkali Containing Bioglass (저농도 알칼리 생체유리의 물성 및 Hydroxyapatite 형성)

  • 김용수;김철영
    • Journal of the Korean Ceramic Society
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    • v.31 no.12
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    • pp.1521-1528
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    • 1994
  • To improve mechanical strength of bioglass, it is considered to use the glass as a coating material to alumina, but the difference in thermal expansion coefficient between two materials is too high to make a good coating. The aim of the present study, therefore, is to find out proper glass composition matching its thermal expansion coefficient to that of alumina without losing biocompatibility. In the present work, various glasses were prepared by substituting B2O3 and CaO for Na2O in the glass system of 55.1%SiO2-2.6%P2O5-20.1%Na2O-13.3%CaO-8.9%CaF2 (in mole%), and the thermal expansion property and reaction property in tris-buffer solution for the resulting glasses were measured. The thermal expansion coefficient of the glass was decreased with the substitution of B2O3 for Na2O, and it became close to that of alumina in the glass in which 8 mole% of CaO was substituted for Na2O. Hydroxyapatite formation was enhanced and silica rich layer thickness was decreased with B2O3 substitution for Na2O. CaO substitution for Na2O didn't deteriorated the hydroxyapatite development.

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Effect of Al2O3 on the Structure and Properties of Bioglass (생체 유리의 구조 및 물성에 미치는 Al2O3의 영향)

  • 노종남;황진명;김철영
    • Journal of the Korean Ceramic Society
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    • v.26 no.6
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    • pp.811-819
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    • 1989
  • There have been many studies on the biological phenomena of Bioglasses, which nay be used as implant materials in human body. However, not many works on the Bioglass compositions have been reported. In the present study, the effect of Al2O3 substitution for SiO2 in Bioglass of Na2O-CaO-P2O5-SiO2 system on its structure and properties was examined. Infrared and Raman spectroscopic studies for the glass structural analysis, differential thermal analysis and X-ray diffraction analysis for crystallization of the glass were perfomed. Several physical properties, such as thermal expansion coefficient, softening point, microhardness and reaction phenomena, were also measured. The major crystalline phase, after heat treatment of the glasses, was Na2Ca2(SiO2)3 and the crystal was transformed into other phase with increased substitution of Al2O3. The added Al2O3 reduced non-bridging oxygen in glass structure and thermal expansion coefficient, but increased glass density, sofening point and microhardness. When the glasses are reacted in Tris-buffer solution, the substituted Al2O3 inhibited the formation of hydroxyapatite on the Bioglas surface, and no hydroxyapatite was formed for the sample which contained more than 6wt.% of Al2O3 even if they were reacted for 600 hours.

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Functional Properties of $\alpha$-Lactalbumin Separated from Bovine Whey (우우 유청으로부터 분리한 $\alpha$-락트알부민의 기능적 특성)

  • 홍윤호
    • Food Science of Animal Resources
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    • v.18 no.1
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    • pp.9-18
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    • 1998
  • This study was performed to obtain a large quantity of $\alpha$-lactalbumin ($\alpha$-LA) from milk by an improved separation and purification method. Functional properties-solubility viscosity, emulsifying activity, foamability surface hydrophobicity and gelation-of the purified $\alpha$-LA were investigated, $\alpha$-LA was purified in a large quantity by DEAE-Sephacel chromatography using 0.15M NaCl in 20mM Tris-HCl buffer(pH 7.2), as an eluent. The yield and purity of the purified $\alpha$-LA were 23.6%, 92.5%, respect-ively. The solubility viscosity and emulsifying activity of the purified $\alpha$-LA were 92.2$\pm$2% 3.46$\pm$0.19 cP and 345$\pm$5.0m^2$-/g respecively. The foamability of $\alpha$ -LA was 762 after 5min whipping which was lower than that of WPC and showed decreasing tendency with whipping time. The relative surface hydrophobicity of the $\alpha$-LA was formed when a 10% $\alpha$-LA solution containing 100mM NaCl and 20 mM $CaCl_2$ was heated at 92$^{\circ}C$for 40min. The $\alpha$-LA gel showed 31.5 as hardness and showed low springiness and cohesiveness.

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Effect of Substitution of MgO for CaO on the Bioglass Structure, Properties and Hydroxyapatite Formation (Bioglass에서 CaO 대신 MgO의 치환첨가에 따른 유리구조, 물성 및 Hydroxyapatite형성)

  • 이호필;김철영
    • Journal of the Korean Ceramic Society
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    • v.27 no.8
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    • pp.979-990
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    • 1990
  • The possible use of bioglass as implant materials is due to its biocompatibility to human body. Even if many animal studies for the bioglasses have been performed, their structures and physical properties are not fully understood. In the present work, several investigations such as Raman spectroscopic analysis, density, thermal expansion coefficient, softening temperature, and refractive index measurement were carried out to find the structures and physical properties of bioglasses, where MgO is substituted for CaO in bioglass composition (46.1%SiO2, 24.4%Na2O, 26.9%CaO, 2.6%P2O5 ; mole%). Hydroxyapatite formation on the glass surface reacted in Tris-buffer solution was also examined. When CaO was replaced by MgO, nonbridging oxygen in glass structuer was diminished but the degree of disorder increased. Thermal expansion and softening properties showed the mixed oxide effect. Hydroxyapatite were formed on the surface of 0~11mole% of MgO containing bioglasses, and the thickness of SiO2-rich layer as well as hydroxyapatite layer were unchanged with MgO content. However, the hydroxyapatite was not formed on the surface of the bioglasses containing over 11 mole percent MgO, even if the glasses were reacted for long period.

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A simple chemical method for conversion of Turritella terebra sea snail into nanobioceramics

  • Sahin, Yesim Muge;Orman, Zeynep;Yucel, Sevil
    • Journal of Ceramic Processing Research
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    • v.19 no.6
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    • pp.492-498
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    • 2018
  • In this study, a sea shell was converted into bioceramic phases at three different sintering temperatures ($450^{\circ}C$, $850^{\circ}C$, $1000^{\circ}C$). Among the obtained bioceramic phases, a valuable ${\beta}-TCP$ was produced via mechanochemical conversion method from sea snail Turritella terebra at $1000^{\circ}C$ sintering temperature. For this reason, only the bioceramic sintered at $1000^{\circ}C$ was concentrated on and FT-IR, SEM/EDX, BET, XRD, ICP-OES analyses were carried out for the complete characterization of ${\beta}-TCP$ phase. Biodegradation test in Tris-buffer solution, bioactivity tests in simulated body fluid (SBF) and cell studies were conducted. Bioactivity test results were promising and high rate of cell viability was observed in MTT assay after 24 hours and 7 days incubation. Results demonstrated that the produced ${\beta}-TCP$ bioceramic is qualified for further consideration and experimentation with its features of pore size and ability to support bone tissue growth and cell proliferation. This study suggests an easy, economic method of nanobioceramic production.

Studies on Various Test Conditions and Application of Test Method for Lipoxygenase-l in Soybean (콩의 Lipoxygenase-1 신속 검정방법 확립)

  • 조준형;김영미;윤홍태;김용호;김용욱;김명애
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.6
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    • pp.739-747
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    • 1997
  • This study was performed to clarify various conditions on the test of lipoxygenase-l and to establish the application of new test method for varietal improvement of soybeans in order to decrease beany flavors. Potassium borate and Tris were used as buffer and O.1M potassium borate solution showed the best result for the lipoxygenase-l test. In the range of pH 8.5~9.0 of the buffer, 2mM linoleic acid as substrate was effective. For color development, 100$\mu$l of two solutions(KI and starch) were added to the half soybean seed, successively. The substrate solution included linoleic acid was stored safely for 10 days at 4$^{\circ}C$ in refrigerator and for 4 days at room temperature. The best result was as follows; the 1ml of substrate solution[0.1M potassium borate(pH 9.0), 0.1% Tween-20, 2mM linoleic acid] was added to the chipped half soybean seed in l.5ml plastic tube, waited for 15 minutes, and 100$\mu$l of color development solutions(5% saturated KI in 15% acetic acid, 1% starch) were added to the tube, successively. After 4 hours, the purple color was observed in the upper phase of the plastic tube in the presence of lipoxygenase-1 and milky color in absence of lipoxygenase-1. The purple color was stable from 4 to 24 hours. There was no interfering effect by lipoxygenase-2 and -3. The plastic tube should be placed in the tube stand without shaking during the lipoxygenase-l test.

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Formation of Zn(II) Complexes of Physiological Buffer Amino alcohols in Aqueous Solution (수용액 중에서 생리학적 완충제 아미노 알코올과 Zn(II) 이온과의 착물 형성)

  • Hong, Kyung-Hee;Chun, Yong-Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.11
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    • pp.7555-7563
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    • 2015
  • The complex formation from Zn(II) ion and 2-(2-hydroxyethylamino)-2-(hydroxymethyl)-1,3-propanediol (Monotris), Bis(2-hydroxyethyl)imino-tris(hydroxymethyl)methane(Bistris) in aqueous solution at $25^{\circ}C$ and at an ionic strength of 0.10 M have been studied potentiometrically. For the Zn(II)-Monotris system, in the Monotris (L) complex $ZnL^{2+}$, one of the hydroxyl oxygen atoms as well as the amine nitrogen of the ligand are coordinated. In basic media, the coordinated hydroxyl group is deprotonated. For the Zn(II)-Bistris system, in the Bistris(L) complex $ZnL^{2+}$, two of the hydroxyl oxygen atoms as well as the amine nitrogen of the ligand are coordinated. In basic media, one of the coordinated hydroxyl groups is deprotonated. In very high basic media, an additional hydroxyl group undergoes deprotonation. The equilibrium constants for the formation of $ZnL^{2+}$, $ZnLH_{-1}{^+}$, $ZnLH_{-2}$, $Zn_2L_2H_{-2}{^{2+}}$ and $Zn_2L_2H_{-3}{^+}$ have been determined.

Separation of Soybean Protein by Free-flow Electrophoresis (자유유동 전기이동법에 의한 대두단백질 분리)

  • 한재갑;류화원
    • KSBB Journal
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    • v.10 no.1
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    • pp.63-70
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    • 1995
  • The effect of operating conditions on separation of soybean proteins in a home-made free-flow electrophoresis apparatus was investigated. Measurement of the pH, conductivity, and UV-absorbance(280 nm) were carried out at each run and the purity of the sample was tested with SDS-PAGE analysis. The soybean extract pretreated with Tris and boric acid was mixed with the amino acids composed of glutamic acid, histidine, arginine, glycine(1 mM each) with glycyl-glycine(2mM) and KCl(1mM). When the cellulose acetate was used as a compartment between the electrode and the buffer solution in the cell, pH distribution in the separation cell varied from 3.0 at the anodic side to 8.0 at the cathodic side and had two inflection point. The applied voltage was from 300V to 1000V and the separation was better at a higher voltage but the voltage was limited by the capability of the cooling system due to Joule heat. The proteins focused near the middle of the channel. From the change of pH and conductivity it was found that the ions in the channel moved out to the electrodes through the membrane. In the case when the concentration of the buffer solution was increased 5 times, proteins were focused at 300V. We could not increase up to the ten times of the concentration since the temperature difference between inlet and outlet was more than $25^{\circ}C$ and denaturation of proteins was expected. When ion-exchange membranes were used U-type pH distribution was set up due to the ionic polarization near the membrane. The commercial ampholytes, instead of the mixed amino acids showed not much improvements in purity of the separated sample.

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