ID

Western Blot

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Western Blot, Western Immunoblotting, Western Blotting

  • Indications
  1. Confirmation of an indeterminate or positive less-specific lab result (e.g. ELISA)
  • Mechanism
  1. Detects proteins and peptides separated by gel electrophoresis and identified with labeled antibodies
  • Technique
  1. Sampled cells are lysed and protein extracted (e.g. from patient blood sample)
  2. Extracted proteins are applied to a gel surface
  3. Electrical current applied to gel matrix separates molecules based on charge and mass
  4. Separated molecules are transferred from gel onto paper or nylon membrane (e.g. nitrocellulose)
  5. Radiolabeled or Enzyme-linked Antibody probes (similar to ELISA) detect specific substances