• 제목/요약/키워드: microplate

검색결과 150건 처리시간 0.026초

In Vitro에서 히스타민 측정 시 ELISA법과 HPLC-MS 분석법의 비교 (Comparison of Methods for Measuring Histamine by ELISA and HPLC-MS Assay In Vitro)

  • 이인희;김유현
    • 대한임상검사과학회지
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    • 제47권4호
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    • pp.306-312
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    • 2015
  • 알러지 반응이 일어나면 histamine이 방출되기 때문에 histamine을 정량 측정함으로써 유발된 알러지의 정도를 확인 할 수 있다. 일반적으로는 항원-항체반응으로 microplate reader를 이용하여 흡광도를 측정하여 정량 한다. 본 연구에서는 histamine 방출량을 측정함에 있어서 일반적인 항원-항체반응과 분석 화학적인 방법으로 HPLC-MS를 이용한 방법을 비교하였다. 세포주는 RBL-2H3를 사용하였고, C48/80으로 자극시켜 알러지를 유발하였다. 유발된 알러지는 ${\beta}$-hexosaminidase의 측정으로 탈 과립을 확인하였으며 실험의 정당성을 위하여 세포독성 능을 확인하였다. Histamine 정량에서 항원-항체반응에 의한 측정의 정량한계는 10.257 ppm이었고, HPLC-MS에 의한 정량한계는 0.020 ppm으로 현저한 차이를 보였다. 알러지 활성 및 항 알러지 실험에 있어서 histamine의 측정은 HPLC-MS를 이용한 분석이 더 정밀하고 정확한 실험인 것을 확인하였다.

Monitoring of Environmental Arsenic by Cultures of the Photosynthetic Bacterial Sensor Illuminated with a Near-Infrared Light Emitting Diode Array

  • Maeda, Isamu;Sakurai, Hirokazu;Yoshida, Kazuyuki;Siddiki, Mohammad Shohel Rana;Shimizu, Tokuo;Fukami, Motohiro;Ueda, Shunsaku
    • Journal of Microbiology and Biotechnology
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    • 제21권12호
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    • pp.1306-1311
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    • 2011
  • Recombinant Rhodopseudomonas palustris, harboring the carotenoid-metabolizing gene crtI (CrtIBS), and whose color changes from greenish yellow to red in response to inorganic As(III), was cultured in transparent microplate wells illuminated with a light emitting diode (LED) array. The cells were seen to grow better under near-infrared light, when compared with cells illuminated with blue or green LEDs. The absorbance ratio of 525 to 425 nm after cultivation for 24 h, which reflects red carotenoid accumulation, increased with an increase in As(III) concentrations. The detection limit of cultures illuminated with near-infrared LED was 5 ${\mu}g$/l, which was equivalent to that of cultures in test tubes illuminated with an incandescent lamp. A near-infrared LED array, in combination with a microplate, enabled the simultaneous handling of multiple cultures, including CrtIBS and a control strain, for normalization by the illumination of those with equal photon flux densities. Thus, the introduction of a near-infrared LED array to the assay is advantageous for the monitoring of arsenic in natural water samples that may contain a number of unknown factors and, therefore, need normalization of the reporter event.

Maroon clownfish, Premnas biaculeatus 자어로부터 분리한 Vibrio ponticus KJS1의 특성 (Characterization of Vibrio ponticus KJS1 Isolated from Larvae of Maroon clownfish, Premnas biaculeatus)

  • 김종수;노섬;강봉조
    • 한국어병학회지
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    • 제20권1호
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    • pp.25-31
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    • 2007
  • 비브리오증은 해수어의 중요한 질병중 하나로 알려져 있다. 본 연구에서 maroon clownfish (Premnas biaculeatus)를 대상으로 해수 관상어 종묘 생산 기술 개발 과정 중 부화 3~7일령 사이에 섭이를 하지 않고 힘없이 유영하면서 폐사하는 개체들로부터 Vibrio 균주를 분리하였다.분리균주의 16S rRNA gene 염기서열 분석결과 Vibrio ponticus CECT5869 와 99.8%의 상동성을 보여 Vibrio ponticus KJS1으로 동정하였다.GN2 microplate (BioLog, USA)를 이용한 생화학 시험결과에서 dextrin 및 sucrose를 포함한 35가지 기질 이용능이 확인되었고, α-cyclodextrin을 비롯한 52가지 기질은 분해하지 못하였으며, glycogen을 비롯한 7가지 기질은 약한 이용능을 나타내었다.분리 균주에 대한 항생제 감수성 실험 결과에서는 oxolinic acid, flumequine, ciprofloxacin, ofloxacin, norfloxacin, nalidixic acid, pefloxacin, oxycycline hydrochloride에 높은 감수성을 나타내었다.

효소면역법에 의한 닭 전염성 후두기관염 바이러스 항체 측정에 관한 연구 (Detection of Antibody to Infectious Laryngotracheitis Virus by Enzyme Linked Immunosorbent Assay)

  • 임숙경;위성하;최정옥;고홍남
    • 한국동물위생학회지
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    • 제15권1호
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    • pp.32-45
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    • 1992
  • In order to establish and enzyme-linked immunosorbent assay to ILTV, field virus strain of ILTV was propagated in chorioallantoic membrane of the embryonated eggs. purified and used as antigen. The antisera selected from the field samples and immunized chickens based on serum neutralization test were used as the standard positive and negative sera in all tests. It was found that optimal antigen concentration was $2{\mu}g$ of protein per well and a 1 : 100 dilution of standard serum showed low background optical density with negative serum and high P/N values of positive sera. A 1 : 500 dilution of the rabbit anti-chicken IgG peroxidase conjugate produced a high P/N values and thirty minutes was chosen as suitable time to read the optical density of the enzyme substrate reaction and optical density was consistent during the 16 hours after stopper was treated. When coated antigen was kept on microplate for varying time up to 16 hours at $4^{\circ}C$ or $37^{\circ}C,$ no significant difference was observed between the treatment. The coated antigen could be kept without change of antigenicity for at least one month at $-70^{\circ}C,\; -20^{\circ}C,\; 4^{\circ}C$ and room temperature. When blocking buffer contanining bovine serum albumin was mixed directly with conjugate and serum at 10% level induced higher P/N values compared to blocking antigen coated microplate with the same blocking buffer. The coefficience of correlation between ELISA and SN test was 0.577. When antibody response of chickens, vaccinated with ILTV, was examined by ELISA and SN test, antibody rising and decay pattern between the two test was similar until 11 weeks of age. However 12 weeks onward antibody titer checked on by SN test was slightly lower than that tested by ELISA.

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Microplate Fixation without Maxillomandibular Fixation in Double Mandibular Fractures

  • Song, Seung Wook;Burm, Jin Sik;Yang, Won Yong;Kang, Sang Yoon
    • 대한두개안면성형외과학회지
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    • 제15권2호
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    • pp.53-58
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    • 2014
  • Background: Maxillomandibular fixation (MMF) is usually used to treat double mandibular fractures. However, advancements in reduction and fixation techniques may allow recovery of the premorbid dental arch and occlusion without the use of MMF. We investigated whether anatomical reduction and microplate fixation without MMF could provide secure immobilization and correct occlusion in double mandibular fractures. Methods: Thirty-four patients with double mandibular fractures were treated with open reduction and internal fixation without MMF. Both fracture sites were surgically treated. For bony fixations, we used microplates with or without wire. After reduction, each fracture site was fixed at two or three points to maintain anatomical alignment of the mandible. Interdental wiring was used to reduce the fracture at the superior border and to enhance stability for 6 weeks. Mouth opening was permitted immediately. Results: No major complications were observed, including infection, plate exposure, non-union, or significant malocclusion. Five patients experienced minor complications, among whom the only one patient experienced a persistant but mild malocclusion with no need for additional management. Conclusion: This study showed that double mandibular fractures correction with two-or three-point fixation without MMF simplified the surgical procedure, increased patient comfort, and reduced complications, due to good stability and excellent adaptation.

우 Casein의 면역효소분석법 (Enzyme-Linked Immuno-Sorbent Assay for Bovine Caseins)

  • 염행철
    • 한국가축번식학회지
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    • 제16권2호
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    • pp.87-102
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    • 1992
  • 본 연구는 우 casein을 신속, 정확하게 분석할 수 있는 특수 면역효소분석법을 개발하였다. Biotin이 연결된 casein과 peroxidase-conjugated avidin을 사용하였으며 면역화시킨 닭의 난황으로부터 추출한 항체를 이용하여 분석하였다. Sulfo-N-hydroxy succinimido biotin을 사용하여 casein에 biotin을 연결시키고 microplate에 고정한 뒤 peroxidase-conjugated avidin을 결합시켰다. 생산된 항체는 $\alpha$-와 $\beta$-casein에 특이적이었으며, 유청단백질, IgG, 우혈청 알부민과 교차반응은 면역효소분석법과 Western blot에서 나타나지 않았다. 본 분석법의 민감도는 2ng에서 20$\mu\textrm{g}$이었으며 Standard와 시료의 분석시 뚜렷한 평행곡선이 형성되었다. Intra-assay와 Inter-assay의 변이계수는 각각 5.5와 5.7%이었다. 그리고 비유 초기의 casein량을 조사한 결과 분만전 3일경부터 급격히 상승하는 것을 알 수 있었다.

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Microplate-Based Oxygen Radical Absorbance Capacity (ORAC) Assay of Hydrophilic and Lipophilic Compartments in Plasma

  • Kwak Ho Kyung;Blumberg Jeffrey B.;Chen Chung Yen;Milbury Paul E.
    • Nutritional Sciences
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    • 제9권1호
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    • pp.48-54
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    • 2006
  • Methods have been developed to evaluate the total antioxidant capacity of foods and plasma but limitations are associated with their ability to determine precisely the contribution of lipophilic antioxidants in a lipid milieu as well as interactions among them Thus, we modified the Oxygen Radical Absorbance Capacity (ORAC) assay to determine the peroxyradical scavenging ability of both hydrophilic and lipophilic compartments in plasma The hydrophilic ORAC assay was performed in a phosphate buffer system utilizing 2,2'-azobis (2-amidinopropane) dihydrochloride as a peroxyradical generator and fluorescein as the target The lipophilic ORAC assay was carried out in a dimethylsulfoxide :butyronitrile (DMSO/BN, 9:1 v/v) system using 2,2'-azobis (2,4-dimethyl valeronitrile) as a peroxyradical generator and BODIPY C11 581/591 as the target Analyses were conducted in bovine serum supplemented with water - and lipid - soluble antioxidants and in human plasma. Albumin (0.5$\sim$5 g/dL) and uric acid (0.1$\sim$0.5 $\mu$mol/L) increased hydrophilic ORAC values in a dose-dependent fashion ($R^{2}$=0.97 and 0.98, respectively) but had no impact on lipophilic ORAC values. $\alpha$-Tocopherol (15$\sim$200 $\mu$mol/L) increased lipophilic ORAC values in a dose-dependent fashion ($R^{2}$=0.94); neither $\alpha$-tocopherol nor $\beta$-carotene had an impact on hydrophilic ORAC values. However, addition of $\beta$-carotene at physiological concentration (0.23$\sim$1.86 $\mu$mol/L), either alone or in combination with other carotenoids, had no significant impact on lipophilic ORAC values. Thus, while assays of 'total antioxidant capacity' in biological matrices would be a useful research and clinical tool, existing methods are limited by the lack of complete responsiveness to the full range of dietary antioxidants.

검정콩 청국장 조제를 위한 Bacillus megaterium SMY-212의 분리 및 균학적 특성 (Isolation and Microbiological Characteristics of Bacillus megaterium SMY-212 for Prepartion of Black Bean Chungkugjang)

  • 손미예;권선화;성찬기;이상원;박석규
    • 생명과학회지
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    • 제11권4호
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    • pp.304-310
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    • 2001
  • In order to produce a high quality and functional black bean Chungkugjang, a bacterium which has potent enzyme activities(protease:124.8 U/$m\ell$, $\alpha$-amylase: 78.2U/$m\ell$, glucoamylase; 13.9U/$m\ell$, ), was isolated by using the halo zone method and identified by morphological, cultural and biochemical properties, and then finally confirmed by GP-microplate identification system as Bacillus megatrium SMY-212. The isolated SMY-212 system was also high in the formation of mucoid material and fibrinolytic activity.

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The Isolation of Acetylcholinesterase Inhibitory Constituents from Lycoris radiata using On-line HPLC-biochemical Detection System

  • Yang, Hee-Jung;Yoon, Kee-Dong;Chin, Young-Won;Kim, Young-Choong;Kim, Jin-Woong
    • Natural Product Sciences
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    • 제16권4호
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    • pp.228-232
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    • 2010
  • Bioactivity-guided fractionation using on-line HPLC biochemical detection system on $CHCl_3$-soluble fraction of Lycoris radiata led to the isolation of deoxylycorenine (1), O-demethylhomolycorine (2), galanthamine (3), lycoramine (4), mixture of $6{\alpha}$-and $6{\beta}$-haemanthidine (5), and lycorine (6), identified by spectroscopic data and physicochemical property. Among the isolated compounds, 1, 3 and 6 showed acetylcholinesterase inhibitiory activities with $IC_{50}$ values of 18.0, 12.0 and $16.6\;{\mu}M$, respectively, in in vitro colorimetric microplate assay.

Sandwich ELISA 방법을 이용한 숙주유래 단백질의 검출

  • 성혜윤;김창민;민홍기;용군호
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.254-254
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    • 1994
  • 현재 국내에서 시판되고 있는 생물공학 의약품매 혼입될수 있는 숙주유래 단백질을 검출하기 위하여 숙주계로 사용되고 있는 Saccaromyces cerevisiae KCTC 1720과 Escherichis coli k12의 total protein을 분리 정제하여 토끼와 guinea pig으로부터 total protein 항체를 얻었다. 이때 토끼항체의 단백질 농도는 yeast의 경우에 4.05mg/m1, E. coli의 경우에 7.14mg/m1이었고, guinea pig의 단백질농도는 yeasat의 경우에 1.90mg/m1이었고 E. coli의 경우에 7.17mg/m1이었다. S. cerevisiae와 E. coli를 숙주로 하여 생산된 생물공학 의약품의 숙주유래 단백질을 검출하기 위하여 guinea pig항체를 96 well microptate에 흡착시키고 검체와 토끼항체의 순으로 microplate에 첨가하는 방법인 sandwich ELISA방법올 사용하였다. 이 방법을 생물공학 의약품의 숙주유래 단백질 검출에 적용한 결과 사람 성장 호르몬의 경우에는 5ng/vial 이하로 검출되었다. 또한 생물학적 제제 생물공학 제품의 경우에는, B형 간염백신제재와 인터페론 감마는 1ng/vial 이하로 검출되었고 인터페론 알파의 경우에는 25ng/vial이하로 검출되었다. 또한 이 방법은 현재 개발되어 시판되고 있는 생물공학 의약품 내에 혼입된 숙주유래 단백질을 검출하는데 쓰일 것이다.

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