• Title/Summary/Keyword: Horseradish Peroxidase(HRP)

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Electrochemical and Spectroscopic Studies on the Interaction between DNA and the Product of Enzyme-catalyzed Reaction of OPD-H2O2-HRP

  • Niu, S.Y.;Zhang, S.S.;Ma, L.B.;Jiao, K.
    • Bulletin of the Korean Chemical Society
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    • v.25 no.6
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    • pp.829-832
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    • 2004
  • The pure product of 2,3-diaminophenazine was prepared by the enzyme-catalyzed reaction of ophenylenediamine-$H_2O_2$-horseradish peroxidase and characterized by UV/Vis spectroscopy, IR spectroscopy and NMR spectroscopy. The electrochemical behaviour of 2,3-diaminophenazine on the glassy carbon electrode was studied. The interaction between 2,3-diaminophenazine and deoxyribonucleic acid was studied by cyclic voltammetry method and UV/Vis spectroscopy, which indicated that the interaction between them is intercalation. The influence of reacting time was also studied. The binding ratio of the 2,3-diaminophenazine-DNA complex is calculated to be 1 : 2 and the binding constant is to be $5.07{\times} 10^3L{\cdot}mol^{-1}$ at room temperature.

Development of Total Cholesterol Detection System by Fluorescence Chromatography (형광 크로마토그래피에 의한 콜레스테롤 측정법의 개발)

  • Oh, Sang-Wook
    • Journal of Food Hygiene and Safety
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    • v.24 no.2
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    • pp.148-153
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    • 2009
  • In this study, we have developed a fluorescence chromatographic assay for the quantification of total cholesterol in serum, which is a well-known risk predictor for cardiovascular diseases. The new assay system consists of a chromatographic strip in a cartridge, enzyme buffer containing cholesterol esterase, cholesterol oxidase, horseradish peroxidase, and color developer AEC, and a laser fluorescence scanner. The correlation coefficient (r) between cholesterol concentration and relative fluorescence units was 0.968 in the new assay, showing a reliable linearity through the tested range of cholesterol. Recovery test and comparability with a Hitachi 747 instrument showed 106.5-94% and r = 0.939 (p<0.001), respectively. The new assay system for cholesterol was developed as a pre-POCT platform conducted in clinics since it is fast (8 min) and uses a small volume of sample ($5\;{\mu}l$), and it may be applied for on-site diagnostics to replace expensive automated biochemical analyzer.

Detection of Food-Grade Hydrogen Peroxide by HRP-Biocomposite Modified Biosensors

  • Chang, Seung-Cheol
    • Journal of Food Hygiene and Safety
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    • v.32 no.6
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    • pp.447-454
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    • 2017
  • A new amperometric biosensor has been developed for the detection of hydrogen peroxide ($H_2O_2$). The sensor was fabricated through the one-step deposition of a biocomposite layer onto a glassy carbon electrode at neutral pH. The biocomposite, as a $H_2O_2$ sensing element, was prepared by the electrochemical deposition of a homogeneous mixture of graphene oxide, aniline, and horseradish peroxidase. The experimental results clearly demonstrated of that the sensor possessed high electrocatalytic activity and responded to $H_2O_2$ with a stable and rapid manners. Scanning electron microscopy, cyclic voltammetry, and amperometry were performed to optimize the characteristics of the sensor and to evaluate its sensing chemistry. The sensor exhibited a linear response to $H_2O_2$ in the range of 10 to $500{\mu}M$ concentrations, and its detection limit was calculated to be $1.3{\mu}M$. The proposed sensing-chemistry strategy and the sensor format were simple, cost-effective, and feasible for analysis of "food-grade $H_2O_2$" in food samples.

Studies on the Rapid and Competitive Enzyme-linked Immunosorbent Assay for the Detection of Thyroxine (T$_4$) in Human Sera (경쟁적 효소면역측정법을 이용한 환자 혈청 내 Thyroxine (T$_4$)의 검출)

  • Sang-Wook Park;Jong-Bae Kim
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.11-15
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    • 1999
  • Thyroxine (3,5,3',5'-L-tetraiodothyronine; T$_4$) is the most commonly measured thyroid hermono for the diagnosis of various thyroid disorders. Although radioimmunoassay (RIA) is still considered as the reference technique for the measurement of T$_4$, it is generally regarded that RIA has its primary disadventages in handling the wastes and controling the human and material resources. Therefore, establishment of enzyme-linked immunosorbent assay (ELISA) has of great significance. To verify the usefulness of our enzyme immunoassay, we have obtained the standard dose response curve of T$_4$ in patient's sera which is inversely proportional to the amount of herseradish peroxidase (HRP) conjugated monoclonal antibody of T$_4$ bound to the wells. The correlation coefficient (r) between the ELISA and chemiluminescent assay was 0.444 (n=38). Thus we have investigated the establishment of rapid and sensitive competitive ELISA assay method for detection of T$_4$ in patient's sera.

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Retrograde Tracer Studies of Tecto-Reticulospinal Pathway and Dorsal Lateral Geniculate Nucleus on GluR1- and GluR4-Immunoreactive Neurons in the Hamster Superior Colliculus (Tecto-reticulospinal pathway (TRS)와 dorsal lateral geniculate nucleus (dLGN)에서 역행성이동추적물질 이용 햄스터 상구에서 GluR1-, GluR4- 면역반응 신경세포 연구)

  • Choi, Jae-Sik;Lee, Jea-Young;Jang, Yu-Jin;Lee, Eun-Shil;Jeon, Chang-Jin
    • Journal of Life Science
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    • v.20 no.1
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    • pp.1-8
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    • 2010
  • We recently reported the distributions of AMPA ($\alpha$-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate) receptor subtypes glutamate receptors (GluR) 1 and GluR4 in the superior colliculi (SC) of hamsters with antibody immunocytochemistry and the effect of enucleation on these distributions. We also compared these labelings to those of calcium-binding proteins calbindin D28K, calretinin, parvalbumin, and GABA. In the present study, we investigated whether the GluR1- and GluR4-immunoreactive (IR) neurons are interneurons or projection neurons by injection of the retrograde tracer horseradish peroxidase (HRP) into one of each major ascending and descending pathways of the SC. HRP injections were made into a tecto-reticulospinal pathway (TRS) and dorsal lateral geniculate nucleus (dLGN). Animals were then allowed to recover and to survive for 48 hr before perfusion. Sections containing retrograde-labeled neurons were then treated for GluR-immunoreactivity. HRP injections proved that only a small population of the GluR1-IR cells project into TRS (1.4%) and dLGN (2.6%). However, a large subpopulation of GluR4-IR cells project into TRS (32.7%). The differential compositions of inter/projection neurons, along with our previous studies on the separate distribution of the GluR subunits, its differential co-localization with calcium-binding proteins and GABA, and differential reactions to enucleations, strongly imply the functional variety of the receptor subunits in visual behavior responses.

Enzymatic Synthesis of Flame Retardant Phenolic Polymers Catalyzed by Horseradish Peroxidase (Horseradish Peroxidase 효소촉매에 의한 난연성 페놀고분자의 합성)

  • Park, Han Sol;Park, Jung Hee;Lee, Hak Sung;Ryu, Keungarp
    • Korean Chemical Engineering Research
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    • v.51 no.1
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    • pp.111-115
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    • 2013
  • The optimum synthetic conditions of poly(p-phenylphenol) by horseradish peroxidase in dioxane:water (80:20 v/v) mixtures were studied. The stability against thermal degradation and structural properties of the synthesized phenolic resins were investigated by thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC), respectively. The synthetic yield of poly(p-phenylphenol) increased upon the increase of the amount of enzyme up to 0.25 mg HRP/mL, then leveled off for further increase of the enzyme usage. When sodium acetate (100 mM, pH 4~6) and sodium phosphate (100 mM, pH 7~9) were used as the buffering salts for the aqueous component (20% v/v), the synthetic yield of the resin increased at higher pH of the aqueous buffer. But when the pHs of the aqueous buffer were 6 and 9, the synthetic yield strongly depended on the types of the buffering salts; if sodium phosphate was used instead of sodium acetate at pH 6, the yield decreased by about 15% and if sodium bicarbonate was used instead of sodium phosphate, the yield decreased by almost 20%. When the pH range of the aqueous buffer was from 4 to 7, the addition of a radical mediator, 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonate) (ABTS), up to 2 mM improved the synthetic yield of the resin by about 10%. TGA experiments revealed that the thermal stability of the resin synthesized in dioxane:water (100 mM sodium phosphate, pH 9) (80:20 v/v) was high having the char yield of 47% upon the heating at $800^{\circ}C$. DCS results showed that the structures of the polymers synthesized in acidic aqueous buffers were different from those of the polymers synthesized in the basic aqueous buffers. However, all the synthesized resins were found to have the property of the thermosetting resins.

MORPHOLOGICAL ANALYSIS OF DEVELOPMENTAL CHANGES IN SOMA AREA OF DIGASTRIC MOTONEURONS IN THE RAT TRIGEMINAL MOTOR NUCLEI (흰쥐의 두힘살근 운동신경세포의 발달과정에 관한 형태학적 분석)

  • Kim, Jae-Hyun;Park, Mi-Hwa;Paik, Sang-Kyoo;Ma, Su-Kyung;Baek, Sang-Heum;Cha, Du-Won
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.2
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    • pp.137-142
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    • 2005
  • To analyze the developmental changes in soma diameters of digastric motoneurons, wheat-germ agglutinin conjugated horseradish peroxidase (WGA-HRP) was injected into the digastric muscle and visualized the retrogradely HRP-labeled motoneurons through tungstate/tetramethylbenzidine (TMB) and following diaminobenzidine (DAB) reactions. The results obtained from Sprague-Dawley rats at postnatal days 1 (P1), 10 (P10) and 30 (P30) indicated as follows: firstly, soma diameters of digastric motoneurons showed unimodal distribution in all postnatal days examined; secondly, the period of P1 to P10 (period 1) showed about 2 times faster growth rate than that of P10 to P30 (period 2); thirdly, the smallest soma examined in each postnatal day exhibited slower growth rate with that of the largest one (increase ratio in soma diameters from P1 to P30, smallest vs. largest = 1.62 : 1.93); Finally, relative growth rates a day showed again that period 1 had faster growth rate than that of period 2. Consequently, developmental changes in soma diameters of digastric motoneurons resulted in very different growth rates between both periods. This implies that the growth of the soma is almost completing within P10 and thereafter growing slowly. The period 1 and 2 are corresponding to sucking and sucking/masticatory period, respectively. Therefore present study providing morphological changes in soma diameters of digastric motoneurons suggests that both periods and their different growth rates of the motoneurons in each period may closely be related with each other.

Fabrication of a influenza A H1N1 sensor using ZnO nanostructure (산화아연 나노구조를 이용한 H1N1 인플루엔자 A 바이러스 센서 제작)

  • Jang, Yun-Seok;Park, Jung-Il;Nam, Yoon-Kyung;Pak, Jung-Ho
    • Proceedings of the KIEE Conference
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    • 2011.07a
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    • pp.1690-1691
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    • 2011
  • 본 논문에서는 H1N1 인플루엔자 A 바이러스(influenza A H1N1 virus) 검출을 위한 산화아연 나노구조(zinc oxide nano structure) 기반의 전기화학적 면역센서를 제작하고 그 특성을 분석하였다. H1N1 인플루엔자 A 바이러스는 빠른 전파 속도 때문에 정확하고 빠른 검출이 필요하다. 먼저, 2 $mm^2$의 표면적을 갖는 패턴된 금 전극 위에 열수방식(hydrothermal method)으로 성장시킨 산화아연 나노구조가 선택적으로 형성되도록 리프트-오프(lift-off) 방법을 사용하였다. 0.01 M phosphate buffered saline(pH 7.4)에서 2 ${\mu}g$/mL 농도의 1차 항체를 정전기력에 의해 산화아연 나노구조에 고정화한 후, 10 pg/mL ~ 5ng/mL 농도의 H1N1 항원을 적용하여 포획 항체에 결합시키고 HRP(horseradish peroxidase) 효소가 결합된 검출 항체를 항원에 결합시키는 샌드위치 ELISA법을 이용하였다. HRP와 반응하는 TMB(3,3', 5,5'-tetramethylbenzidine)와 과산화수소가 포함된 acetate buffered 용액(pH 5)을 전해질로 사용하고 순환전압전류 측정법(cyclic voltammetry)으로 센서의 특성을 분석하였다. 측정된 순환전압전류그래프(cyclic voltammogram)에서 H1N1 항원 농도 10 pg/mL ~ 5 ng/mL의 응답 전류는 276.47 ${\pm}$ 21.72 nA (평균 ${\pm}$ 표준편차, n=4) ~ 478.89 ${\pm}$ 6.21 nA로 측정되었고, logarithmic하게 증가하는 응답 전류 특성을 보였다.

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Morphological Analysis of the Myelinated Parent Axons that Innervate Rat Upper Molar Pulps in the Trigeminal Ganglion

  • Paik, Sang Kyoo;Kim, Jong Ho;Kim, Tae Heon;Bae, Yong Chul
    • International Journal of Oral Biology
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    • v.40 no.4
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    • pp.175-182
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    • 2015
  • Previous studies suggested that myelinated axons innervating rat molar pulps undergo morphological changes in their peripheral course. However, little information is available on the morphological feature of the parent axons at the site of origin. We therefore investigated the size of the myelinated parent axons and their morphological features at the proximal sensory root of the trigeminal ganglion by horseradish peroxidase (HRP) injection into rat upper molar pulps and subsequent light and electron microscopy. A total of 248 HRP-labeled myelinated axons investigated were highly variable in the size. Fiber area, fiber diameter, axon area (axoplasm area), axon diameter (axoplasm diameter), and myelin thickness were $11.32{\pm}8.36{\mu}m^2(0.80{\sim}53.17{\mu}m^2)$, $3.99{\pm}1.53{\mu}m(1.08{\sim}9.26{\mu}m)$, $8.70{\pm}6.30{\mu}m^2(0.70{\sim}41.83{\mu}m^2)$, $3.13{\pm}1.13{\mu}m(0.94{\sim}7.20{\mu}m)$ and $0.43{\pm}0.23{\mu}m(0.07{\sim}1.06{\mu}m)$, respectively. The g-ratio (axon diameter / fiber diameter) of the labeled axons was $0.79{\pm}0.05$ (0.61~0.91). Axon diameter was highly correlated with myelin thickness (correlation coefficients, r=0.83) but little correlated with g-ratio (r=-0.33) of individual myelinated parent axons. These results indicate that myelin thickness of the myelinated parent axons innervating rat molar pulps increase with increasing axon diameter, thus maintaining a constant g-ratio.

Central Projections of Sensory Information Produced by Topical Application of Capsaicin to the Tongue in the Cat

  • Hong, Seung-Kil;Nahm, Sook-Hyun;Han, Hee-Chul;Suh, Young-Suk;Won, Ran;Lee, Bae-Hwan
    • The Korean Journal of Physiology
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    • v.25 no.1
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    • pp.87-99
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    • 1991
  • In order to elucidate whether capsaicin applied topically acts as a pain-producing subastance or as a tastant, neuronal activities of the chorda tympani nerve(CN), lingual nerve(LN), solitary tract nucleus(STN), and trigeminal nucleus(TGN) were recorded while thermal and taste stimuli, and capsaicin were being applied topically, and algesics intra-arterially to the tongue of cats anesthetized with ${\alpha}-chloralsoe$. In addition, the STN neurons were examined after wheat germ agglutinin-horseradish peroxidase(WGA-HRP) was applied to the CN. The CN fibers responded to taste and thermal stimuli, algesics, and capsaicin. Responses to capsaicin were significantly correlated with those to taste and thermal stimuli. The LN fibers mainly responded to mechanical and thermal stimuli, algesics, and capsaicin. Responses to capsaicin were significantly correlated with those to algesics. The STN neurons responded to taste and thermal stiumli, algesics, and capsaicin. Responses to capsaicin were significantly correlated with those to taste and thermal stiumli in somewhat different fashion from those of the CN fibers. The TGN neurons mainly responded to mechanical stimuli, algesics, and capsaicin. Correlations between responses to capsaicin and any others were not significant. After WGA-HRP was applied to the CN, the STN neurons which receive input from the CN were identified largely in the medio-ventral portion to the solitary tract. These results suggest that capsaicin produce taste as well as pain sensation. Sensory information evoked by capsaicin can be conveyed to the STN, especially medio-ventral portion, via the CN as gustatory information on the one hand, and to the STN or TGN via the LN as noxious information on the other. In addiation, the noxious information may be conveyed to the STN via the CN.

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