蛇王淘金20.pptVIP

Stan Hitomi Coordinator – Math Science San Ramon Valley Unified School District Danville, CA Kirk Brown Lead Instructor, Edward Teller Education Center Science Chair, Tracy High School and Delta College, Tracy, CA Sherri Andrews, Ph.D. Curriculum and Training Specialist Bio-Rad Laboratories Essy Levy, M.Sc. Curriculum and Training Specialist Bio-Rad Laboratories Why TeachSize Exclusion Chromatography? Size Exclusion Chromatography Size Exclusion Chromatography Instructors Powerful teaching tool Laboratory extensions Real-world connections Link to careers and industry Standards based Size Exclusion Chromatography Kit Advantages Standards Based Can be used in Biology, Chemistry, or Physical Science Sufficient materials for 8 student work stations Easy preparation Easy visualization of separation Can be completed in one 45 minute lab session Study how the structure and biochemical properties of molecules are related to their separation Workshop Time Line * * Introduction Comparison of different types of column chromatography Separation of a mixture of biomolecules by size exclusion chromatography Types of Column Chromatography Affinity Hydrophobic Interaction (HIC) Ion Exchange Anion Cation Gel Filtration or Size Exclusion (SEC) * * Affinity Chromatography Uses an affinity tag - allows molecules to bind to the column - specific to the tagged protein of interest - Examples: HIS-Tag, antibody, GST-Tag Proteins not bound pass through the column A buffer is used to elute the protein from the column * * /Molecular%20Biology%20Glossary.files/image047.gif Ion Exchange Chromatography Beads in the column are charged Anion - positively (+) charged beads Cation- negatively (-) charged beads Molecule to be purified will have the opposite charge from the beads in the column Molecules not binding to the beads pass through the column A counter-charged buffer is used to elute the molecule of interest /Molecular%20Biology%