BCH210H1 Lecture Notes - Lecture 11: Sodium Chloride, Organic Compound, Unified Atomic Mass Unit

• School: UTSG
• Department: Biochemistry
• Course: BCH210H1
• Professor: Stavroula Andreopoulos

Document Summary

Lecture 11 outline: analysis of proteins, chromatography methods, sequencing methods. Analysis of proteins: purification by chromatography, amino acid composition, amino acid sequence. Protein purification: characterization of protein structure and function requires a pure sample, protein samples obtained from cell lysates or by recombinant dna methods are usually mixtures, methods are needed to purify proteins for analysis. Types of chromatography: gel filtration (size-exclusion) chromatography, affinity chromatography, hplc: high pressure liquid chromatography (reverse phase) Gel filtration: gel filtration separates by size and shape (globular is faster than asymmetrical, falling through beads, affinity chromatography uses glucose, hplc exploits hydrophobicity of proteins. Separates on size (mw) and shape: chromatography column separates protein by size (mw, column matrix (resin) consists of porous beads with varying pore size, calibrate column with protein of known size (see when it elutes) Smaller proteins are selectively trapped (and slowed), because they enter the pores, enter the space through the bead.