Lecture6-8ColorSlides

Lecture6-8ColorSlides - Interfering Substances of the...

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Unformatted text preview: Interfering Substances of the Protein Assay: SDS Soap structure resembles the blue form of dye. Low [SDS] can interfere with binding of dye to protein. SDS v. Dye for protein. High [SDS] forms micelles. Nonpolar Fuid core. Stabilizes green form. SDS Micelles v. Protein for dye. [SDS] > the critical micelle concentration (CMC) results in saturation of green form. Protein has virtually no chance. Detergents in Protein Sample - -- Water Surrounding Nonpolar Regions - Hydrophobicity Water must stagger itself to reduce polarity and thus reduce repulsion with the nonpolar regions of a molecule. The hydrogen bonds between water are very weak due to conFguration and also cannot form multiple layers as water that surrounds an ion. Only requires a small amount of thermal energy to disrupt thin layer of water. Nonpolar groups have higher probable collisions with each other. When nonpolar groups come together, there is less repulsion because of reduce surface area exposure to the polar solvent.- ------- ------ ------ LAB PRACTICAL PRACTICE: Tues. 7/6: 1:00 - 9:00 pm Check-in Lab 1077 Young Last Check-in time is 7:00 pm. No Lab Coat, No goggles, No Shirt, No Shoes, No Service! Practice duplicates of two 1:10 samples for dye binding protein and LDH enzyme assays. See VOH for this protocol. Standards will NOT be run during practice. Children Under 12 get in free! * * Michael J. Davies, Photochemical and Photobiological Sciences, 2004 3(1), 17-25 Oxidation Rate of oxidation is slow. Peroxidation leads to increased degradation. Chance of loss of Enzyme function >>> lack of protein dye-binding. Proteases Rate of proteolytic digestion fast Proteases released and activated by lysed cells (similar to apoptosis) Chance of loss of enzyme function > lack of protein dye-binding. But lack of protein binding can be signiFcant. * x F B = m s ! 2 x F f = v f F c = m m !...
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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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Lecture6-8ColorSlides - Interfering Substances of the...

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