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Isoelectric Focusing definitions

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  • Isoelectric Focusing
    Electrophoresis technique that separates proteins by their isoelectric points using a stable pH gradient in a gel.
  • Electrophoresis
    Method that uses an electric field to separate charged biomolecules, such as proteins, based on their properties.
  • Isoelectric Point
    Specific pH at which a protein's net charge is zero, causing it to stop migrating in an electric field.
  • pH Gradient
    Continuous range of pH values established in a gel, decreasing linearly from high to low, crucial for protein separation.
  • Protein Band
    Distinct region in the gel where a protein accumulates after migration stops at its isoelectric point.
  • Net Charge
    Overall electrical charge of a protein, determined by the ionization state of its amino acid groups at a given pH.
  • Ionization
    Process by which amino acid groups gain or lose protons, altering the charge of a protein as pH changes.
  • Ionizable Group
    Functional part of a protein that can accept or donate protons, affecting the protein's charge in different pH environments.
  • pKa
    Value indicating the pH at which an ionizable group is half protonated, influencing protein charge during migration.
  • Immobile Gradient
    Stable pH distribution in the gel that remains unchanged during the separation process, ensuring accurate protein focusing.
  • Electric Field
    Force applied across the gel to drive protein migration based on their charge until reaching their isoelectric point.
  • Gel
    Medium used in isoelectric focusing to establish a pH gradient and facilitate protein separation.
  • Migration
    Movement of proteins through the gel under the influence of an electric field, halted at their isoelectric point.
  • Sample Loading
    Initial placement of protein mixture at the top of the gel before applying the electric field for separation.
  • Linear Gradient
    Type of pH gradient where values decrease evenly from top to bottom of the gel, essential for precise protein separation.