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Biochemical Separation Techniques
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Gel Electrophoresis
Principle: Utilizes an electric field to migrate charged molecules through a gel matrix. Use-Case: Separating DNA or protein fragments based on size for analysis.
Mass Spectrometry
Principle: Ions are separated based on their mass-to-charge ratio. Use-Case: Identifying and quantifying molecules, such as in metabolomics.
Isoelectric Focusing
Principle: Separates proteins based on their isoelectric point (pI) in a pH gradient. Use-Case: Analyzing protein charge properties and purification.
Western Blotting
Principle: Combines gel electrophoresis and antibody-based detection. Use-Case: Detecting specific proteins in a sample after their separation by size.
Size Exclusion Chromatography
Principle: Molecules are separated by size through a gel filtration matrix. Use-Case: Separating proteins or polymers based on their hydrodynamic volume.
Dialysis
Principle: Molecules are separated based on size using a semi-permeable membrane. Use-Case: Removing small molecules or ions from proteins or nucleic acids in solution.
Centrifugation
Principle: Uses centrifugal force to separate components in a mixture based on density differences. Use-Case: Separating cellular components such as nuclei, mitochondria from a cell lysate.
Two-Dimensional Gel Electrophoresis
Principle: Combines isoelectric focusing and SDS-PAGE to separate proteins by charge and size. Use-Case: Comprehensive analysis of complex protein mixtures, like whole cell lysates.
Chromatography
Principle: Separates mixture components based on differential affinities to stationary and mobile phases. Use-Case: Purifying proteins or small molecules, such as in drug manufacturing.
Affinity Chromatography
Principle: Separates molecules based on specific binding interactions with a ligand. Use-Case: Purifying a specific protein from a complex mixture using an antibody or substrate.
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