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Lecture 16

Lecture 16 con't - BIOL308.pdf

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Department
Biology
Course
BIOL 308
Professor
Bruce Wolff
Semester
Fall

Description
Lecture 16 contMeasuring gene expression by measuring translationSodium dodecyl sulfate polyacrylamide gel electrophoresis SDS PAGE and western blottingSDS negatively charged detergent binds to hydrophobic protein regions helps protein unfoldingProteins bind lots of SDS in constant ratio to their mass SDS gives them a negative charge proportional to their massequal charge density per unit length protein intrinsic charge is maskedElectrophoresis under denaturing conditions proteins migrate towards the positive electrode when voltage is applied to a gelSeparate on the basis of molecular weight rather than intrinsic chargeProteins detected by different stains gel stained with coomassie brilliant blue silver etcWestern BlottingFirst step is SDS PAGEproteins are denatured by heat and detergent SDS and electrophoresed separating on the basis of sizeThe proteinsare transferred to a membrane western blottingSimilar to DNA in southern and northernThe membrane is incubated with an antib
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