웨스턴 블럿
- 최초 등록일
- 2010.08.02
- 최종 저작일
- 2009.05
- 6페이지/ 한컴오피스
- 가격 3,000원
* 본 문서는 한글 2005 이상 버전에서 작성된 문서입니다.
한글 2002 이하 프로그램에서는 열어볼 수 없으니, 한글 뷰어프로그램(한글 2005 이상)을 설치하신 후 확인해주시기 바랍니다.
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웨스턴 블럿 실험결과 보고서
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본문내용
1) Western blot
Western blot, also called immunoblot, uses antibody to identify a specific protein of interest. Similar to ELISA, the nitrocellulose membrane that contains the resolved proteins will be first incubated with an unrelated protein to block nonspecific protein binding sites on the membrane, and then blotted with a primary antibody that is specific for the protein of interest. The unbound primary antibody will be washed away. The solution used for washing contains a detergent and a high concentration of salt that eliminate nonspecific binding of the antibody to the nitrocellulose through hydrophobic or ionic interaction. Next is the addition of a secondary antibody that is specific for the primary antibody and conjugated to horseradish peroxidase (HRP). After washing, a substrate of HRP will be added. HRP catalyzes a reaction that transforms the colorless substrate into a colored product, which visualizes the protein band.
Gel electrophoresis/Western blot is one of the powerful methods for detecting a particular protein in a complex mixture. This technique combines the superior resolving power of gel electrophoresis, the specificity of antibodies, and the sensitivity of enzyme assays. Compared to gel electrophoresis/Western blot, ELISA is better for the quantification rather than identification of a protein of interest.
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