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Western Blot

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Topic updated on 03/21/15 7:24pm

Overview
  • Also called Immunoblot 
  • Purpose
    • Test to identify proteins
      • primarily used as a confirmatory test for HIV infection due to high specificity (near 100%)
  • Process
    • protein separated via gel electrophoresis
    • antigen blotted onto nitrocellulose paper
    • serum of patient added
    • anti-human immunoglobulin antibody added
      • conjugated to color producing enzyme or radiolabel
    • test determined to be reactive if patient serum reacts to two of three HIV gene products
    • HIV gene products tested for by western blot: 
      • p24 ( capsid protein)
      • gp 160 envelope protein that is cleaved to form gp 41 and gp 120
        • gp41 (envelop protein that mediates fusion and entry)
        • gp 120 (envelop protein that mediates attachment to CD 4 cells)
 

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