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Primary+Structure+of+Proteins-B (1)

Primary+Structure+of+Proteins-B (1) - Electrophoresis 1...

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Effects of Salts on Protein Solubility Increase in solubility Increase in salt concentration “salting in” protein aggregation by ionic interactions salt ions screen charges in protein
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Effects of Salts on Protein Solubility Increase in solubility Increase in salt concentration “salting out” Salt ions withdraw hydration water from protein molecules
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Fractionation of Proteins by “Salting Out” Stepwise increase in (NH 4 ) 2 SO 4 concentration supernatant supernatant
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Ion-Exchange Chromatography (IEC) H N 3 CO 2 H N 3 CO 2 H N 3 C O 2 H N 3 C O 2 H N 3 CO 2 H N 3 C O 2 C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + C + Cation exchange matrix
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Size Exclusion Chromatography (SEC) Synonyms: Gel Permeation Chromatography, Gel Filtration Chromatography
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Affinity Chromatography (AC)
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SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE)
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SDS-PAGE
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Two Dimensional (2D) Gel
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Unformatted text preview: Electrophoresis 1. Dimension: Isolelectric focusing (IEF) 2. Dimension: SDS-PAGE Radioactively labeled total proteins from E. coli Summary: Protein Purification Overview of Protein Sequencing Overview of Protein Sequencing Cleavage of Disulfide Bonds Site-specific Cleavage of Peptide Bonds by Proteases (Endopeptidases) Cleavage of Peptide Bonds at Met-residues by Cyanogen Bromide N-terminal Sequencing by Edman Degradation (OH-) (anhydrous F 3 C-CO 2 H) (H + ) After each step separation by RP chromatography Cycle 1 Cycle 2 Cycle 3 Cycle 4 N-terminal Sequencing by Edman Degradation Mass Spectrometry Electrospray Ionisation (ESI) Apomyoglobin Mass = 16,952.2 Tandem Mass Spectrometry...
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