PP09 - 1.Ide ntify ingPro te insinC rude Extrac ts by We...

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12/16/09 1 1. Identifying Proteins in C rude Extracts  by Western Blotting 2. Protein Bioinform atics
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12/16/09 2 Things To Learn From This Week’s Lab: • Antibodies are used to identify specific proteins on Western blots, and specific components of cells. THE FINAL PART OF THE PROTEIN WORK: • The Western blotting Technique. Bioinformatics-definition -analyses of protein sequence using database -prediction of subcellular localization -prediction of protein function -prediction of domain structure and function
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12/16/09 3 Electroblotting Voltage-driven transfer of proteins from  a  gel to a transfer m em brane SDS  Gel Transfer  Membrane Electro- blotting     Gel    after  blotting Transfer  Membrane After  Blotting *
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12/16/09 4 Western Blotting:   Detection of a  Specific Protein Using Protein- Specific Antibodies Also called an   im m unoblot ”.
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12/16/09 5 Antibodies are Proteins that are  produced by an anim al im m une  system Each antibody   recognizes   and   binds   to one  specific   target   m olecule. The target m olecule is called an  antigen .
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12/16/09 6 Antibodies are Y -Shaped Proteins.  Each antibody has two  binding sites for Antigens “C onstant region” is identical  for all antibodies m ade in the  sam e species of organism . Antigen Binding Site Antigen Binding Site “Variable region” (antigen  binding site) is different  for different antibodies. 
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12/16/09 7 Each Antibody can Recognize and  Bind to a  Single Type of Protein  (antigen)  in a C rude Extract of  Thousands of Proteins. proteins
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12/16/09 8 Antibodies Bind to Epitopes on an Antigen Protein Antigen epitope epitope Epitopes on a protein are usually just a few amino acids in size Antibody
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12/16/09 9 Polyclonal Antibodies Are Raised in anim als such as  Rabbits  or  Goats  for Research Purposes. The anim al is injected with a  purified protein .  Over tim e, the anim al develops a high titer of  antibodies  that recognize and bind to the injected  protein.
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This note was uploaded on 10/06/2010 for the course BIO 205L taught by Professor Hanson during the Fall '07 term at University of Texas at Austin.

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PP09 - 1.Ide ntify ingPro te insinC rude Extrac ts by We...

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