1. Separation of Proteins by Gel 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