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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– Read about different methods of protein
separation– Never use any methods while in school or in
teaching
What do we know about protein structure?
• Building blocks?• Primary structure?
• Secondary structure?
• Tertiary Structure?
• Quaternary Structure?
What makes one Protein Different from another?
There are two properties of proteins used to separate them
• Size or MW
• Electric Charge
This lab will use Agarose gel electrophoresis to separate proteins
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
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?
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