Gel Electrophoresis - PowerPoint PPT Presentation

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Gel Electrophoresis

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Gel Electrophoresis Steps Breakdown the protein into amino acids Break amide/peptide bond Called hydrolysis (opposite of condensation) Acid or base and heat required ... – PowerPoint PPT presentation

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Title: Gel Electrophoresis


1
Gel Electrophoresis
2
Steps
  • Breakdown the protein into amino acids
  • Break amide/peptide bond
  • Called hydrolysis (opposite of condensation)
  • Acid or base and heat required
  • Add protein to gel containing a buffer
  • Apply a voltage
  • Amino acids move based on mass and charge
  • Dye amino acids
  • Ninhydrin
  • Measure distance traveled,
  • Compare to known

3
Set-up
4
Why amino acids move
  • Amino acids separate based on their isoelectric
    point and molar mass
  • Isoelectric point
  • This is the pH where they net charge of amine and
    carboxylic acid groups cancel out
  • NH3-C-COO-

5
  • pH of buffer is more basic than isoelectric
    point, than amino acid will have a negative
    charge and move toward postive electrode
  • There are fewer hydrogen atoms around to
    protonate it
  • NH2-C-COO-
  • pH of buffer is more acidic than isoelectric
    point, than amino acid will have a positive
    charge and move to negative electrode
  • More H atoms around to protonate
  • NH3-C-COOH
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