PROTEIN PURIFICATION &
CHARACTERIZATION
April 11 - 24, 2012
Application Deadline: February 17, 2012
Instructors:
Richard Burgess, University
of Wisconsin, Madison
Albert Courey, University of California, Los
Angeles
Sue-Hwa Lin, University
of Texas M.D.Anderson Cancer Center
Michael Marr,
Brandeis University
This
course is for scientists who are not familiar with techniques of protein
isolation and characterization. It is a rigorous program that includes
laboratory work all day and a lecture with discussion session every
evening. Each student will become familiar with each of the major
techniques in protein purification by actually performing four separate
isolations including: (i) a regulatory protein
from muscle tissue: (ii) a sequence-specific, DNA binding protein: (iii)
a recombinant protein overexpressed in E. coli; and (iv) a membrane-bound
receptor. A variety of bulk fractionation, electrophoretic, and
chromatographic techniques will include: precipitation by salts, pH, and
ionic polymers; ion exchange, gel filtration, hydrophobic interaction,
and reverse phase chromatography; lectin
affinity, ligand affinity, oligonucleotide affinity, and immunoaffinity chromatography; polyacrylamide gel
electrophoresis, and electroblotting; and high
performance liquid chromatography. Procedures will be present for
solubilizing proteins from inclusion bodies and refolding them into
active monomeric forms. Methods of protein characterization will be
utilized to include immunological and biochemical assays, peptide
mapping, amino acid analysis, protein sequencing, and mass spectrometry.
Emphasis will be placed on strategies of protein purification and
characterization. Guest lecturers will discuss protein structure,
modification of proteins, methodologies for protein purification and
characterization, and applications of protein biochemistry to cell and
molecular biology.
This
course is supported with funds provided by the National Cancer Institute
Cost
(including board and lodging): $3,535
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