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Lecture5 - MCB 110L Spring 2011 Lecture 5 Qing Zhong 306...

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Unformatted text preview: MCB 110L Spring 2011 Lecture 5 2/2/2011 Qing Zhong 306 Barker Hall Assays, assays, assays …a biochemist’s jeweled tools!!! How much total protein is present? How much of this protein is the kinesin Cin8p? Desired features of any assay for ANYTHING: Simple Quick Accurate Reproducible Sensitive Specific Enzyme assay I-Continuous assays • Spectrophotometric- also called colorimetric assays if color is changed. Fluorometric-Fluoescence is when a molecule emits light of one wavelength after absorbing light of a different wavelength. Calorimetric-heat absorbance Chemiluminescent-emission of light by a chemical reaction. ECL, luciferase. Enzyme assay I-Discontinuous assays • Radiometric » Radiometric assays measure the incorporation of radioactivity into substrates or its release from substrates. • Chromatographic » Measures product formation by separating the reaction mixture into its components by chromatography Factors to control in assays • Salt concentration › High salt kills (1-500mM) • Effect of temperature ›...
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Lecture5 - MCB 110L Spring 2011 Lecture 5 Qing Zhong 306...

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