Lecture12Summer10BWHandouts-1

Lecture12Summer10BWHandouts-1 - 1 Drying the column •...

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Unformatted text preview: 1 Drying the column • Beads shrink and creates large cracks in the bead layer. • Lowers binding and increases velocity of all AMP-binding proteins. Packing Problems • Degassing – Air pockets. More pockets like drying out column. • Displacing the top of the bed layer. – Uneven entry into the bed layer. – DeFnes zone of migration. Chromatography: Zones of Migration A + B B A c c 2 Chromatography: Zones of Migration A + B B A MORE ZONE BROADENING Multipath Broadening 1 2 Longitudinal Diffusion Zone Broadening • Diffusion – Should not stop mobile phase. – Just the right drip rate – Not too fast – less change to bind. • Bead sizes are uniform – Reduce random migration • While Group A checks the 60% sup and pellet for enzyme… • Group B the afFnity column • 1. Check empty column is not clogged • 2. Add buffer to column; Volume = bead layer. (EB says 1 ml) • 3. Draw line to help gauge when beads are packed....
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This note was uploaded on 10/07/2010 for the course POLI SCI 40 taught by Professor Sch during the Spring '10 term at UCLA.

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Lecture12Summer10BWHandouts-1 - 1 Drying the column •...

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