TLS Online TPP Program

#Id: 2813

#Unit 13. Methods in Biology

Reversed-phase liquid chromatography differs from most other forms of chromatography in that the stationary phase is essentially inert and only non-polar (hydrophobic) interactions are possible with analytes.

TLS Online TPP Program

#Id: 2814

#Unit 13. Methods in Biology

Proteins differ in the amount of hydrophobic amino acids that are present on their surface. This difference can be exploited in salt fractionation but can also be used in a higher resolution method using hydrophobic interaction chromatography (HIC).

TLS Online TPP Program

#Id: 2815

#Unit 13. Methods in Biology

HIC column is made up of phenyl-Sepharose, where phenyl groups are bonded to the insoluble support Sepharose.

TLS Online TPP Program

#Id: 2816

#Unit 13. Methods in Biology

When in the protein solution divalent salts, or multivalent salts the protein separated in form of aggregate.

TLS Online TPP Program

#Id: 2817

#Unit 13. Methods in Biology

In the salt fractionation most polar will be fractionated with large amount of salt.

TLS Online TPP Program

#Id: 2818

#Unit 13. Methods in Biology

In the HIC some highly hydrophobic proteins may not even be eluted in the total absence of salt in such case water-miscible organic solvent such as propanol or ethylene glycol to the column buffer solution.