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Lecture6-8ColorSlides

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

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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 fluid 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.
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Detergents in Protein Sample -
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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 configuration 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.
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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!
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