Handout 3 - Technology - Outline Technology...

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Outline Technology  Protein purification  Antibodies  Immunoflorescence  GFP  Transformation, Transfection, Transgenics  X-ray crystalography
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Strategies to study proteins involved in a process : Biochemistry : Purify the proteins involved and study their behavior  in vitro.  Recapitulate the process in a test tube. Identify a minimum set  of proteins that can make something happen. Genetics : Isolate a mutant that is defective in the process.  Also  engineer organisms. Cell   Biology : Watch the process in a microscope.  Follow the dynamics  of particular proteins in a living cell.  Does inhibiting the function of a  specific protein in a living cell block the process you are interested in? None of these fields are independent:  Cell biologists do biochemistry, Cell biologists use genetics, Geneticists  use cell biology….
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Basic Techniques for  Studying Biology at the Cell  and Molecular Level “Biochemistry”                        Protein How do we purify a protein of interest?   
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Fraction A Fraction B Fraction C Distinct Subsets of Proteins Fractionation - Separate by Physical & Chemical properties Cell Extract 1000’s of proteins
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Column Chromatography
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Standard Column Chromatography Protein Elution via  [Salt] in Buffer
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How do you know which  fraction contains the  protein you are interested  in?
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