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Antibody Affinity Purification

HP Huan Pang

Published: May 5, 2012 DOI: 10.21769/BioProtoc.208 Views: 14440

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Abstract

Antibody purification is performed to concentrate and enrich antigen-specific antibodies and lower the background signal during detection by removing any non-specific proteins. Affinity purification makes use of specific binding interactions between molecules, and is very simple and efficient.

Materials and Reagents

  1. PBS
  2. NaCl
  3. KCl
  4. Na2HPO4
  5. KH2PO4
  6. Glycine
  7. HCl
  8. Tris-HCl
  9. BSA
  10. Azide
  11. Affinity resin
  12. Sepharose
  13. 10x PBS (see Recipes)
  14. 1x PBS + 1 M NaCl (see Recipes)
  15. 0.1 M glycine-HCl (pH 2.8) (see Recipes)
  16. 1 M Tris- HCl (pH 8.5) (see Recipes)

Equipment

  1. Centrifuges
  2. Affinity resin
  3. Spectrometer

Procedure

Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.

Category

Biochemistry > Protein > Isolation and purification

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