How Does pH Affect Peptide Separation in Electrophoresis?

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SUMMARY

The optimal pH for electrophoresis separation of peptides with isoelectric points (pI) of 10.3 and 7 is determined by their charge at various pH levels. At pH 8, the peptide with a pI of 10.3 carries a positive charge while the peptide with a pI of 7 carries a negative charge, facilitating effective separation. The key to successful electrophoresis is ensuring that the peptides have opposite charges, which enhances their migration towards the respective electrodes. Therefore, pH 8 is the recommended choice for achieving optimal separation.

PREREQUISITES
  • Understanding of isoelectric points (pI) and their significance in peptide behavior.
  • Knowledge of electrophoresis techniques and their operational principles.
  • Familiarity with pH effects on protein charge and migration.
  • Basic grasp of peptide chemistry and structure.
NEXT STEPS
  • Research the impact of pH on protein charge and stability in electrophoresis.
  • Explore different electrophoresis techniques, such as SDS-PAGE and native PAGE.
  • Learn about buffer systems used in electrophoresis and their pH optimization.
  • Investigate the role of temperature and ionic strength in peptide migration during electrophoresis.
USEFUL FOR

Biochemists, molecular biologists, and laboratory technicians involved in protein analysis and purification through electrophoresis techniques.

Melow
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Hi all, I have a question about this exercise:

Which is the best option of pH for electrophoresis separation of two peptides with isoelectric points of 10.3 and 7 respectively? The given options of pH are 3,8 or 10.

I've thought that if pH is more than 7, then, there will be positive charge for the peptide of 10.3 pI, and negative charge for the peptide of 7 pI, but i don't know how to manage th sign of the charges to get the best eletrophoresis separation. Is it better to have both of them with the same charge or the opposite charge?

<Moved to homework section. No template>
 
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How does the sign of the charge influence how the peptides migrate during electrophoresis?
 

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