COMM 1076 Lecture Notes - Lecture 10: Polyvinylidene Fluoride, Bio-Rad Laboratories, Powdered Milk
Document Summary
The purpose of this experiment is to determine the presence of specific protein that contains a very specific amino acid sequence -hhhhhh (i. e. , a his-tag). This experiment takes two weeks and introduces a wide variety of laboratory techniques along the way. The lab begins by teaching the process of fractionation. The protein assay is followed by the introduction of gel electrophoresis(sds-page) and the western blot. In gel electrophoresis, an applied voltage separates the proteins by molecular weight. The proteins are then transferred to a membrane where they can be treated with antibodies to identify protein of interest. All instructions were followed as the protocol handout available on: blackboard, in biochem lab. Ii section, under course document tab, file name western blot : materials, supplies and equipment. All the reagents for this experiment were provided by teaching assistants except for 1x sdspafe gel running buffer: 4-20% sds-page gel, pre-stained protein mw ladder number 7.
