Name: monitoring protein adsorption with solid-state nanopores
Uploaded: 2011-12-02T13:00:00
Description: Watch this Scientific Journal Video about Monitoring Protein Adsorption with Solid-state Nanopores at JoVE.com

저자들은 조언 존 Grazul (코넬 대학), 안드레 Marziali (밴쿠버에서 브리티시 컬럼비아 대학)와 빈센트 토바드 - Cossa을 (오타와 대학) 감사드립니다. 이 작품은 미국 국립 과학 재단 (DMR - 0706517와 DMR - 1006332)와 국립 보건원 (R01 - GM088403)에서 부여로 일부 자금이다. 재료 연구 과학 및 공학 센터 (MRSEC) 프로그램 (DMR 0520404) - nanopore 드릴링은 국립 과학 재단 (National Science Foundation)의 지원으로 재료 연구 코넬 센터 (CCMR)의 전자 현미경 시설에서 수행되었다. 실리콘 질화물 점막의 준비는 국립 과학 재단 (부여 항법 - 0335765)에서 지원하는 코넬 NanoScale 시설, 국립 나노기술 인프라 네트워크의 일원에서 실시되었습니다.

1. 실리콘 질화물 점막에서 고체 nanopores의 제조 <ol><li> 200 kV의 가속 전압에 황비홍 Tecnai F20 S / TEM을 가져와. 다른 S / TEM을 사용하는 경우, 가속 전압보다 크거나 200 kV 9이어야합니다 </li><li> TEM 샘플 홀더에 20 nm의 두꺼운 SPI 실리콘 질화물 창 그리드를로드하고 소유자로부터 오염 물질을 제거하는 30 초 동안 산소 플라즈마로 닦아주십시오. </li><li> S / TEM에 샘플을로드하고 아래 펌프로 진?...

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고체 표면에 27-29로 단백질의 자연 흡착는 biochip 응용 프로그램 및 기능 하이브리드의 biomaterials의 새로운 클래스의 디자인 분야의 숫자에 근본적으로 중요합니다. 이전 연구는 고체 표면에 adsorbed 단백질이 측면이나 이동성을 크게 탈착 속도를 표시하지 않는 것으로 나타났습니다, 따라서 단백질 흡착은 일반적으로 돌이킬 수와 특이 현상이 프로세스를 30-32로 간주됩니다. 고체 ?...

<table border="1"><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> Tecnai F20 S / TEM </td><td> 페이 </td><td></td><td> S / TEM은 가속 전압 ≥ 200kV 및 현장 방출 소스를 필요로합니다. </td></tr><tr><td> TEM 20 NM 두꺼운 실리콘 질화 막 창 </td><td> SPI </td><td> 4163SN - BA </td><td></td></tr><tr><td> 축삭 Axopatch 200B 패치 - 클램프 증폭기 </td><td> 분자 장치 </td><td></td><td></td></tr><tr><td> 축삭 Digidata 1440A </td><td> 분자 장치 </td><td></td><td></td></tr><tr><td> 10 소프트웨어를 pCLAMP </td><td> 분자 장치 </td><td></td><td> 전기 생리학 데이터 수집 및 분석 소프트웨어 </td></tr><tr><td> 황산 </td><td> Fisher 과학 </td><td> A300 </td><td></td></tr><tr><td> 과산화수소 </td><td> 피셔 과학 </td><td> H325 </td><td></td></tr><tr><td> 실리콘 O - 링 </td><td> McMaster - 카 </td><td> 003 S70 </td><td> 또한 PDMS를 사용하여 </td></tr><tr><td> 실버 와이어 </td><td> 시그마 - 올드 리치 </td><td> 348,759 </td><td> 전극에 대한 </td></tr><tr><td> SPC 기술, D 하위 연락처, 핀 </td><td> 뉴어크 </td><td> 9K4978 </td><td> 전극에 대한 </td></tr><tr><td> 염화칼륨 </td><td> 시그마 </td><td> P9541 </td><td></td></tr><tr><td> 인산 칼륨 이염 </td><td> 시그마 </td><td> P2222 </td><td></td></tr><tr><td> 인산 칼륨 일염기의 </td><td> 시그마 </td><td> P5379 </td><td></td></tr><tr><td> PDMS </td><td> 다우 코닝 </td><td></td><td> Sylgar 184 Elastomer가 설정할 수 있습니다. 챔버를 만들기위한. </td></tr><tr><td> Kwik - 캐스트 밀봉제 </td><td> 세계 정밀 계측기 </td><td> KWIK - CAST </td><td> 빠른 행동 실리콘 밀봉제 </td></tr><tr><td> 핫 플레이트 </td><td> 피셔 과학 </td><td></td><td></td></tr><tr><td> 패러데이 케이지 </td><td></td><td></td><td></td></tr></table>

monitoring protein adsorption with solid-state nanopores

고체 nanopores은 지역 구조와 유연성 핵산 1-6, 단백질 7 전개, 그리고 다른 리간드 8 바인딩 선호도를 조사하는 단일 분자 수준에서 측정을 수행하는 데 사용되었습니다. 9-12 저항 펄스 기술로 이러한 nanopores을 결합함으로써, 그러한 측정은 다양한 조건 하에서와 라벨 3에 대한 필요없이 할 수 있습니다. 저항 펄스 기술에서는 이온 소금 솔루션은 nanopore의 양쪽에 도입입니다. 따라서, 이온은 지속적으로 현재의 결과, 적용 transmembrane 잠재력에 의해 챔버의 한쪽에서 다른 구동됩니다. nanopore에 analyte의 파티션은 단일 분자 정보를 추출하는 분석할 수이 현재에 잘 정의된 편향을 발생합니다. 이 기법을 사용하여 nanopore 벽에 단일 단백질의 흡착은 다양한 범위의 아래에 모니터할 수 있습니다조건 13. microfluidic 장치의 크기 축소로, 하나의 단백질과 함께 이러한 시스템의 상호 작용이 우려되고 있기 때문에 단백질 흡착은 중요성이 높아지고있다. 이 프로토콜은 쉽게 nanopore 드릴링 의무가 다른 박막, 또는 작용 질화 표면에 확장될 수 있습니다 질화물 필름에 바인딩 단백질에 대한 신속한 분석을 설명합니다. 단백질의 다양한 솔루션 및 denaturing 조건 하에서 광범위한 탐험 수 있습니다. 또한이 프로토콜은 nanopore 분광법을 사용하여보다 기본적인 문제를 탐구하는 데 사용할 수 있습니다.

Watch this Scientific Journal Video about Monitoring Protein Adsorption with Solid-state Nanopores at JoVE.com

Monitoring Protein Adsorption with Solid-state Nanopores

무기 표면에 단백질이 아닌 구체적인 흡착을 모니터링하는 고체 nanopores를 사용하는 방법을 설명합니다. 방법은 실시간과 단일 분자 수준에서 탐색할 수 흡착 수있게 저항 펄스의 원칙을 사용합니다. 하나의 단백질 흡착 과정 멀리 평형에서이기 때문에, 우리는 양적 단백질 흡착의 상수 명백한 처음 주문 반응 속도의 결정뿐만 아니라 및 Langmuir 흡착 상수 사용, 합성 nanopores의 병렬 배열의 고용을 제안합니다.

고체 Nanopores와 모니터링 단백질 흡착

Solid-state nanopores have been used to perform measurements at the single-molecule level to examine the local structure and flexibility of nucleic acids ...

Research

JoVE Journal

Bioengineering

Registered Bioimaging of Nanomaterials for Diagnostic and Therapeutic Monitoring

진단 및 치료 모니터링을위한 Nanomaterials 등록 Bioimaging

Nanomedications can be carried by blood borne monocyte-macrophages into the reticuloendothelial system (RES; spleen, liver, lymph nodes) and to end ...

연구

생체공학

Fluorescence detection methods for microfluidic droplet platforms

microfluidic 비말 플랫폼 형광 검출 방법

The development of microfluidic platforms for performing chemistry and biology has in large part been driven by a range of potential benefits that ...

Monitoring the Wall Mechanics During Stent Deployment in a Vessel

선박의 스텐 트를 배포하는 동안 벽 역학 모니터링

Clinical trials have reported different restenosis rates for various stent designs1. It is speculated that stent-induced strain concentrations on the ...

Mass Cytometry: Protocol for Daily Tuning and Running Cell Samples on a CyTOF Mass Cytometer

대량 세포 계측법 : CyTOF 질량 Cytometer에 매일 조정하고 실행 세포 샘플에 대한 프로토콜

In recent years, the rapid analysis of single cells has commonly been performed using flow cytometry and fluorescently-labeled antibodies. However, the ...

Multi-analyte Biochip (MAB) Based on All-solid-state Ion-selective Electrodes (ASSISE) for Physiological Research

Multi-analyte Biochip MAB Based on All-solid-state Ion-selective Electrodes ASSISE for Physiological Research

생리 연구를위한 모든 고체 이온 선택성 전극 (ASSISE)를 기반으로 다중 분석 바이오칩 (MAB)

Lab-on-a-chip (LOC) applications in environmental, biomedical, agricultural, biological, and spaceflight research require an ion-selective electrode (ISE) ...

Multi-Scale Modification of Metallic Implants With Pore Gradients, Polyelectrolytes and Their Indirect Monitoring In vivo

Multi-Scale Modification of Metallic Implants With Pore Gradients, Polyelectrolytes and Their Indirect Monitoring In vivo

모공 그라디언트, 고분자 전해질 및 그들의 간접적 인 모니터링과 금속 임플란트의 멀티 스케일 변경<em&gt; 생체 내</em

Metallic implants, especially titanium implants, are widely used in  clinical applications. Tissue in-growth and integration to these implants in  the ...

Construction and Setup of a Bench-scale Algal Photosynthetic Bioreactor with Temperature, Light, and pH Monitoring for Kinetic Growth Tests

키네 성장 시험을위한 온도, 빛 및 pH 모니터링을 갖춘 벤치 규모의 조류 광합성 생물 반응기의 건설 및 설치

The optimal design and operation of photosynthetic bioreactors (PBRs) for microalgal cultivation is essential for improving the environmental and economic ...

In Vesiculo Synthesis of Peptide Membrane Precursors for Autonomous Vesicle Growth

자율 적인 소포 성장을 위한 펩티드 멤브레인 전구체의 베시큘로 합성

Compartmentalization of biochemical reactions is a central aspect of synthetic cells. For this purpose, peptide-based reaction compartments serve as an ...

Functional Transcranial Doppler Ultrasound for Monitoring Cerebral Blood Flow

대뇌 혈류 를 모니터링하기위한 기능성 경두개 도플러 초음파

Functional transcranial Doppler ultrasound (fTCD) is the use of transcranial Doppler ultrasound (TCD) to study neural activation occurring during stimuli ...

Labeling of Extracellular Vesicles for Monitoring Migration and Uptake in Cartilage Explants

연골 내의 마이그레이션 및 섭취 모니터링을 위한 세포외 소포 라벨링

Extracellular vesicles (EVs) are used in different studies to prove their potential as a cell-free treatment due to their cargo derived from their ...