연세대학교 2016년 2학기 A+ 받은 리포트입니다.
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2. Experimental Background
In this investigation, the main objective is to understand the principle as well as step-by-step procedures of performing SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and western blot to analyze proteins.
SDS-PAGE is used for separation of DNA and especially for separation of proteins based on their molecular weight via polyacrylamide gel by applying an electrical field. This technique involves two different discontinuous gels known as stacking and resolving gels.
According to figure 1, the position (size) of a 3rd marker which is ovalbumin (46.379kDa) serves as a standard size marker to roughly predict and determine the site of α-tubulin. Here, α-tubulin must be in between bovine serum albumin (79.236kDa) and closer to the 3rd marker since the molecular weight of α-tubulin is 55kDa.
Basic Molecular Biotechnology Laboratory Booklet Department of Biotech. Yonsei University.
By Analyzing Location and Intensity of the Specific Reaction, Expression Details of the Target Proteins in the given Cells or Tissue Homogenate Could Be Obtained. Western Blotting Could Detect Target Protein Which Is as Low as 1ng Due to High Resolution O. "Western Blot Principle." Western Blot Principle. Web. 07 Nov. 2016. <https://www.bosterbio.com/protocolandtroubleshooting/western-blot-principle>.
"Introduction to PAGE." Sigma-Aldrich. Web. 07 Nov. 2016. <http://www.sigmaaldrich.com/technical-documents/articles/biology/sdspage.html>.
"SDS-PAGE." SDS-PAGE. Web. 07 Nov. 2016. <http://www.assayprotocol.com/molecular-biology/electrophoresis/denaturing-page>.