BCH210H1 Lecture Notes - Lecture 8: Polyacrylamide Gel Electrophoresis, Sodium Dodecyl Sulfate, Subtilisin

• School: UTSG
• Department: Biochemistry
• Course: BCH210H1
• Professor: Stavroula Andreopoulos

Document Summary

Sds-page: sodium dodecyl sulfate - polyacrylamide gel electrophoresis. Sds: denaturing detergent and dissolves membranes and solubilizes proteins. Small molecules will be located towards the bottom of the gel: larger at the top. Making sure the proteins is made and is the correct size. Transfer onto nitrocellulose (paper-like) (a) represents single polypeptides. Epitope tag: sequence of amino acids inserted into or onto protein, can be recognized by a specific antibody. Lane 1: the lysate (aka sonicate) from e. coli - contains all proteins. Lane 2: the flow-through from the ni-affinity column: things that don"t bind. Lane 3: the purified expressed (his-tagged) fusion protein eluted from the column with. Lanes 1", 2" and 3": same as lanes 1, 2, and 3, but blotted with an anti-fusion protein antibody. Secreted in large amounts from many bacillus species. X-ray structures have been determined (globular protein, several a-helices, b-sheet) 2 domains of 1 polypeptide: active site in the middle to hydrolyse.