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Biochemical Separation Techniques

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Gel Electrophoresis

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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.

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Mass Spectrometry

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Principle: Ions are separated based on their mass-to-charge ratio. Use-Case: Identifying and quantifying molecules, such as in metabolomics.

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Isoelectric Focusing

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Principle: Separates proteins based on their isoelectric point (pI) in a pH gradient. Use-Case: Analyzing protein charge properties and purification.

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Western Blotting

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Principle: Combines gel electrophoresis and antibody-based detection. Use-Case: Detecting specific proteins in a sample after their separation by size.

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Size Exclusion Chromatography

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Principle: Molecules are separated by size through a gel filtration matrix. Use-Case: Separating proteins or polymers based on their hydrodynamic volume.

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Dialysis

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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.

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Centrifugation

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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.

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Two-Dimensional Gel Electrophoresis

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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.

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Chromatography

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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.

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Affinity Chromatography

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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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