연세대학교 2016년 2학기 A+ 받은 리포트입니다.
실험관련 topic에 대해 많은 조사를 했기에 이 자료 하나로 충분할거에요.
2. Experimental Background
5. Results & Discussion
6. Preparation of Western Blot Sample [Calculation]
1. RIPA buffer
Radio-immunoprecipitation assay (RIPA) buffer causes cell wall to break leading to efficient cell lysis and protein solubilization from cultured animal cells. It also avoids interfering with immunoreactivity of proteins, degradation of protein and biological activity. In addition, RIPA buffer is appropriate for immunoprecipitation and western blot assays because it minimizes binding of non-specific protein which results in low background in such assays.
<Results & Discussion>
In conclusion, the main objective of the experiment was to determine the appropriate protein concentration based on Bradford assay. This is to ultimately prepare 75μg/50μL (or 1.5μg/μL) of western blot sample prior to perform western blot assay. In this investigation, due to possible errors and limitations, the results of OD600nm measurement & protein concentration might not be very accurate.
Basic Molecular Biotechnology Laboratory Booklet – Department of Biotech. Yonsei University.
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"Bio-Rad Quick Start Bradford Protein Assay." 4110065 Rev A. Quick Start Bradford Protein Assay. Bio-Rad Laboratories, Inc. Web. 30 Oct. 2016. <http://www.biorad.com/webroot/web/pdf/lsr/literature/4110065A.pdf>.
"R0278 SIGMA RIPA Buffer." RIPA Buffer Safety & Documentation. SIGMAALDRICH, Web. 30 Oct. 2016.<http://www.sigmaaldrich.com/catalog/product/sigma/r0278?lang=ko®ion=KR>