5.AffinityChromatographyfortheIsolationofLDH

5.AffinityChromatographyfortheIsolationofLDH - Affinity...

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Affinity Chromatography for the Isolation of LDH Week 5 Lab S117 Fall 2011
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Overview Post-Lab discussion Standard curves: uses and limitations Comparison of two assays for [protein] Pre-Lab Discussion Chromatography Enzyme Assays
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Standard Curves Assumption: Standard has same response as unknown Limit of detection, limit of linearity, and extrapolation Overly-weighted points Dropping last point gives y = 0.99x + 0.02
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Comparison of Protein Assays Don’t uncritically choose experimental techniques Don’t uncritically accept others’ experimental techniques Strengths and weaknesses of assays Ease and convenience Sensitivity Accuracy and precision
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Pre-lab: Affinity Chromatography Major goals of biochemical research Find biochemical activity Separate compounds with activity Study activity This week Separation with chromatography Confirm activity with assay
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Chromatography Stationary phase Paper Polymer resin Mobile phase (eluent) Gas Liquid Movement Capillary (up)
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5.AffinityChromatographyfortheIsolationofLDH - Affinity...

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