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Separation of Proteins by Gel Electrophoresis

Separation of Proteins by Gel Electrophoresis. Carolina Biological Kit #68-9800. Presented by. Bryan Smith. Why this Kit?. Just Curious! Done chemical tests for proteins Done enzyme activity labs Done Gel Electrophoresis to analyze DNA by separating molecules w/different #Bp

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Separation of Proteins by Gel Electrophoresis

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  1. Separation of Proteins byGel Electrophoresis Carolina Biological Kit #68-9800

  2. Presented by Bryan Smith

  3. Why this Kit? • Just Curious! • Done chemical tests for proteins • Done enzyme activity labs • Done Gel Electrophoresis to analyze DNA by separating molecules w/different #Bp • Read about different methods of protein separation • Never use any methods while in school or in teaching

  4. What do we know about protein structure? • Building blocks? • Primary structure? • Secondary structure? • Tertiary Structure? • Quaternary Structure?

  5. What makes one Protein Different from another?

  6. There are two properties of proteins used to separate them • Size or MW • Electric Charge

  7. This lab will use Agarose gel electrophoresis to separate proteins

  8. Lab particulars • Our protein sample contains two pre-stained proteins both w/overall negative charge • Albumin • Hemoglobin • Work in pairs • Gel is 1% agarose • Class will run two gels

  9. Procedures • Pour two gels using largest well comb • After solidifying, remove comb and orient so wells are on “negative” end • Fill chamber with TBE buffer • Each pair will load one 40uL sample of protein using micropipette • Voltage?

  10. Go over discussion questions How many different scenarios can you think of where separating proteins is essential to a biologist? XXXX – isolating insulin from pancreatic cells

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