• Journal of the Optical Society of Korea
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• 제20권3호
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• pp.431-434
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• 2016

Tris-HCl buffer solution is extensively used in biochemistry and molecular biology to maintain a stable pH for biomolecules such as nucleic acids and proteins. Here we report on the high-precision THz dielectric spectroscopy of a 10 mM Tris-HCl buffer. Using a double Debye model, including conductivity of ionic species, we measured the complex dielectric functions of Tris-HCl buffer. The fast relaxation time of water molecules in Tris-HCl buffer is ~20% longer than that in pure water while the slow relaxation time changes little. This means that the reorientation dynamics of Tris-HCl buffer with such a low Tris concentration is quite different from that of pure water.

• Archives of Pharmacal Research
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• 제4권1호
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• pp.33-41
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• 1981

The prepared dextran sulfates were characterized by measuring the reduced viscosisty at five different concentrations to obtain an intrinsic viscosity in both phosphate and tris buffers, pH 7.4, ionic strength of 0.1 Dextran sulfates having 0.81, 1.06 sulfate groups per hexose unit have reduced viscosity value below 40 ml/g whereas dextran sulfates having 1.21, 1.43, 1.69 sulfate groups per hexose unit have reduced viscosity value over 40 ml/g. Dextran sulfate having 1.21 sulfate groups per hexose unit had highest value of reduced viscosity. The reduced viscosity of dextran sulfate in tris buffer was always higher than that in phosphate buffer regardless of the sulfate content of dextran sulfate. The influence of the sulfation of the dextran sulfate. The influence of the sulfation of the dextran sulfate molecule on the dextran sulfate-LDL interaction was studied with three different dextran sulfate molecules. Dextran sulfate molecules having more than one sulfate group per hexose unit. The dextran sulfate having 0.81 sulfate groups per hexose unit showed considerably different precipitation curves in phosphate and tris buffers. This peculiar behavior of dextran sulfate having 0.81 sulfate groups per hexose unit in the two buffer systems was not noticed with dextran sulfate having more than one noticed with dextran sulfate having more than one sulfate group per hexose unit.

• 한국수정란이식학회지
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• 제17권2호
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• pp.137-143
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• 2002

본 연구는 개의 동결정액 제조 시 동결보호 희석액 내에 첨가되는 당의 종류와 조합이 동결융해 후 정자의 생존율 및 운동성에 미치는 영향과 정자동결 시 straw size가 생존율에 미치는 영향에 대하여 조사하고자 실시하였다. 희석액은 Tris-citric acid extender (Tris-buffer)의 기본용액에 20% Egg-yolk, 8% glycerol, 1% Equex STM paste등을 첨가하였으며, 당 성분으로는 monosaccharide(fructose 및 xylose) 및 disaccharide(trehalose)로 구분하여 최종 70 mM의 농도로 첨가 이용하였다. 본 연구에서는 control (fructose, xylose, trehalose), two combination (Fru＋Tre, Fru＋Xy1, Tre＋Xy1) 및 three combination (Fru＋Tre＋Xy1)으로 구분하여 Tris-buffer에 첨가하였다. Fru＋Tre＋Xy1처리구에서 CASA분석 후 운동성이 fructose, trehalose, xylose, Fru＋Tre, Fru＋Xy1, Tre＋Xy1 처리구에 비해 가장 높았다 (69% vs 58, 61, 50, 65, 20, 54%). 또한 전진운동율은 Fru＋Tre 처리구가 fructose, trehalose, xylose, Fru＋Xy1, Tre＋Xy1, Fru＋Tre＋Xy1 처리구에 비해 가장 높았다 (59% vs 47, 55, 42, 13, 49, 44%). 이러한 결과를 바탕으로 Tris-buffer에 F겨＋Tre를 첨가하여 실험을 수행하였다. 예비동결 10분에서 0.25$m\ell$ straw를 이용하였을 때 10분 예비동결에 0.5 $m\ell$, 5분 예비동결에 0.25 및 0.5 $m\ell$ straw 처리구보다 유의적으로 높은 생존율을 얻었다 (80＋0.0 vs. 65＋7, 68＋16, 58＋8%). 본 연구결과 70 mM Fru＋Tre (two combination)가 첨가구가 가장 높은 전진운동율을 얻었으며, 0.25 $m\ell$ straw에 10분간 예비동결을 실시하는 것이 가장 높은 동결융해 후 생존율을 얻을 수 있었다.

• KSBB Journal
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• 제16권6호
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• pp.533-537
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• 2001

유청 단백질 중에서 $\alpha$-lactalbumin, $\beta$-lactoglobulin를 고성능 막 크로마코그래피를 이용하여 분리하는 것이다. 분리 메카니즘은 음이온 교환작용이며 고정상은 CIM DEAE, QA, So$_3$ disk (16$\times$3 mm)을 사용하였다. 이동상은 buffer A (20 mM Tris-HCI, pH 7.3)와 buffer B (buffer A + 1 M NaCl)를 사용하였으며 $\alpha$-lactalbumin, $\beta$-lactoglobulin를 분리하기 위해서 구배용매 조성법을 사용하였다. 각 이동상의 조성에 따른 최적의 이동상 조성(Buffer A/Buffer B=100/0 - 30/70 vol%, gradient time 1 min, 30/70 - 10/90 vol%, gradient time 2 min)을 실험적으로 얻었고 4 ml/min의 이동상 유속에서 3분내에 $\alpha$-lactalbumin, $\beta$-lactoglobulin를 분리 할 수 있었다. 유청 단백질 중에 $\alpha$-lactalbumin, $\beta$-lactoglobulin을 HPMC을 적용하여 분리하였고, 유청 단백질의 기능적 성질, 분리 방법에 대해 알아보았다.

• Journal of Plant Biology
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• 제36권3호
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• pp.259-266
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• 1993

In Tris-iso-butanol (TIB; Tris buffer pH 8.8 and 1% iso-butanol)-treated chloroplasts, oxygen evolving activity was more inhibited than Tris-treated chloroplasts, but restored highly by 2,6-dichlorophenol-indophenol (DCPIP) and photoreactivation. To understand the mechanism of this results of TIB in photosynthetic electron transport, system, oxygen consumption and evolution of PS I and PS II were measured and protein of the chloroplasts was analysed. In Tris- and TIB-treated chloroplasts, oxygen evolving activity was increased according to the light intensity. Under 48 W·m-2 light intensity, the oxygen evolving activity in both chloroplasts were similar but as the light intensity was increased, TIB-treated chloroplasts showed higher activity. Under 240 W·m-2 light intensity, TIB-treated chloroplasts showed about 25% higher oxygen evolving activity than Tris-treated chloroplasts. Oxygen evolving activity was increased after photoreactivation in both Tris-treated and TIB-treated chloroplasts. Addition of NH4Cl increased the activity in both chloroplasts but in TIB-treated chloroplasts the increase was 30% higher than that in Tris-treated chloroplasts. In PS I, oxygen evolving activity was not inhibited by both treatments whereas in PS II, significant difference was observed between two treatments. Addition of Mn2+ and Ca2+ enhanced oxygen evolution in both Tris- and TIB-treated chloroplasts. Though enhancement was higher in TIB-treated chloroplasts. No difference was observed n protein analysis of the two thylakoid membrane.

• Food Science and Biotechnology
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• 제17권3호
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• pp.651-654
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• 2008

In order to enhance the efficacy of norovirus detection by reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR, this study developed a norovirus mRNA concentration method using poly oligo dT-conjugated magnetic beads. An efficient norovirus detection protocol was performed on commercial ham using 2 viral elution buffers (glycine buffer and Tris beef extract buffer) and 2 concentration solutions [polyethylene glycol (PEG) and zirconium hydroxide]. The different approaches were verified by RT-PCR and nested PCR. This method was performed on ham in less than 8 hr by artificial inoculation of serial dilutions of the virus ranging from 1,000 to 1 RT-PCR unit/mL. The viral extraction and concentration method had 10-fold higher sensitivity using the combination of Tris beef extract buffer and PEG as compared to glycine buffer and zirconium hydroxide. This method proved that RT-PCR and nested PCR have the sensitive ability to detect norovirus in commercial ham, in that norovirus was successfully detected in artificially contaminated samples at a detection level as low as 1-10 RT-PCR unit/mL. Overall, such a detection limit suggests this protocol is both quick and efficient in terms of its potential use for detecting norovirus in meat products.

• The Korean Journal of Physiology
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• 제25권2호
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• pp.115-123
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• 1991

Molecular association of glucose transporters with the other proteins in the plasma membrane was assessed by gel electrophoresis and immunoblot techniques. Approximately $31.5{\pm}5.1%$ of GLUT-4, $64.8{\pm}2.7%$ of clathrin, 48.7% of total protein in the plasma membrane (PM) were found insoluble upon extraction with 1% Tx-100. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the Tx-100 insoluble PM fraction contained about 4 major polypeptides with apparent molecular weight of above 200, 100-120, 80 and 30-35 KDa that were readily removed upon wash with a high pH buffer which is known to remove clathrin and 0.5 M Tris-buffer which is known to remove assembly proteins (AP). Immunoblotting of GLUT4 and clathrin against specific antibodies showed that GLUT-4 and clathrin were co-solubilized up to 84.6% and 82.7% respectively by wash with a high pH buffer and 1% Tx-100. When the membrane was pre-washed with a high pH buffer and 0.5 M Tris solution, GLUT4 and clathrin were not solubilized further suggesting that GLUT4 molecules are in molecular association with clathrin, AP and/or other extrinsic membrane proteins in plasma membrane and the formation of clathrin-coated structures might be involved in insulin stimulated glucose transporter translocation mechanism.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.90-90
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• 2002

The purpose of this study was to investigate the effect of kind and combination of sugars on the viability and acrosome damage of post-thaw spermatozoa in canine. The extender used was Tris-citric acid extender (Tris-buffer) supplemented with 20% Egg-yolk, 8% glycerol, 1% Equex STM paste, and 70 mM sugars such as monosaccharide (fructose and xylose) and disaccharide(trehalose). To evaluate of sugar combination, the sugars supplemented in Tris-buffer were combined such as single (fructose, xylose, trehalose), two combinations (Fruc+Tre, Fruc+xyl, Tre+xyl) and three combinations (Fruc+Tre+Xyl). The concentration of sperm collected were adjusted of 50${\times}$10$\^$6/ per straw for freezing. The frozen spermatozoa were thawed at 37$^{\circ}C$ for 1 min and then analysis for CASA program in Livestock Improvement Main Center, NACF. The motility of post-thaw spermatozoa in Fruc+Tre was higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (79％ vs. 63, 66, 70, 71, 74 and 75%). The progressive motility after CASA analysis in Fuc+Tre group was also higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (67% vs. 53, 57, 60, 61, 62 and 64%). The acrosome damage of post-thaw spermatozoa stained was not significantly different among treatment groups such as fructose, trehalose, xylose, Fru+Tre, Fru+xyl, Tre+xyl and Freu+tre+xyl (17.7, 18.3, 28.0, 17.0, 19.7, 20.0 and 19.0%). The results indicated that the motility and progressive motility of post-thaw spermatozoa in Fru+Tre group was better, and acrosome normality was not different among all groups. The use of Tris-buffer supplemented with Fru+Tre as sugar for frezing of canine spermatosoa could be better and apply to semen banking and artificial insemination.

• Asian-Australasian Journal of Animal Sciences
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• 제16권2호
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• pp.165-171
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• 2003

Sixty semen ejaculates collected at weekly interval from four Murrah Buffalo bulls over a period of seven months (Nov.1999 to May 2000) were used in the present study. Three buffer medium (sodium citrate, TES and Tris) were used for soaking of sephadex. Three grades of sephadex (G-15,G-100, and G-200) were used for preparation of columns. Columns of three different height (one, two and three cm) were used for separation of semen. Twenty semen ejaculates were used in each project. In the first experiment each semen ejaculates was divided into four parts. One part was kept as control and other three parts were passed thought one cm column of sephadex G-15 prepared in three different buffers. There was significant (p<0.05) increase in percent progressive sperm motility and percent live spermatozoa and decrease in percent abnormal spermatozoa and percent spermatozoa with damaged acrosome as well as sperm numbers after filtration through all the three columns. Sperm quality obtained in the filtrate of column prepared in Tris buffer was better in comparison to other two buffers. So the Tris buffer was used in the second trial. Twenty semen ejaculates were used in this experiment. Each semen ejaculate was divided into four parts. One part was kept as control (non-filtered) and other three parts were passed through columns of different grade of sephadex (G-15, G-100 and G-200). Progressive sperm motility and live sperm percentage improved significantly while decline in percent abnormal spermatozoa and percent spermatozoa with damaged acrosome and sperm concentration was observed after filtration through all the columns as compared to control (non-filtered) semen. Since post filtration quality of semen was better in the sephadex G-100 column, therefore it was selected for the next experiment. In third experiment, Tris buffer and sephadex G-100 were used for preparing columns of different height (one, two and three cm) and twenty semen ejaculates were filtered. The quality characteristics of semen (percent progressive sperm motility, percent live spermatozoa and sperm concentration) after filtration through one cm column were significantly (p<0.05) higher than after filtration through columns of two and three cm height. However non -significant (p>0.05) difference due to height of columns was observed for percent abnormal and percent damaged acrosome but 1 cm column comparatively gave better result than of 2 and 3 cm column height.

• Applied Microscopy
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• 제19권2호
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• pp.74-84
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• 1989

인삼 종자의 배유조직에서 Tris 완충액 가용성 저장단백질을 추출한 후 전기 영동적 분석으로 분리하여 $SP_{1}$(MW=160,000)과 $SP_2$(MW=70,000)의 두가지 저장단백질을 정제하였다. 이 두가지 저장단백질을 항원으로 사용하여 토끼에 피하주사하여 항체를 얻었으며, 이 항체를 이용하여 면역 세포화학적 금입자표지법을 실시한 결과, $SP_1$과 $SP_2$ 모두 구형의 protein body내에 산재하여 있음을 확인되었으며, globoid에는 이러한 두가지 단백질중 어느 것도 함유되어 있지 않는 것으로 나타났다. 또한 각각의 protein body에 함유된 $SP_1$과 $SP_2$의 상대적 함량에는 서로 차이가 있음이 확인되었다.