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Rabbit IgG Conjugation to Dynabeads

Xiyan Li Xiyan Li

Published: May 5, 2011 DOI: 10.21769/BioProtoc.63 Views: 16989

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Abstract

This method couples rabbit IgG (or any other proteins serve as affinity reagent) to the surface of magnetic beads (Brand name: Dynabeads). The amine and thiol groups of amino acid residues on the protein are covalently linked with the epoxy group on Dynabeads. The coupled IgG beads can be stored at 4°C for 6-12 month with no obvious loss in reactivity.

Keywords: Dynabeads search Immunoglobulin G search Conjugation search Immunoprecipitation search Affinity purification search

Materials and Reagents

  1. Invitrogen Conjulation kit (143.11D), contains C1, C2, HB, LB, SB solutions
  2. IgG (Sigma-Aldrich, catalog number: I5006-10 mg )
  3. Dynabeads M-270 Epoxy 300 mg (Life Technologies, Invitrogen™)
  4. Phosphate buffered saline (PBS) tablet
  5. 30% NaN3 (fresh)
  6. 20% Tween 20

Equipment

  1. Hula mixer (Life Technologies, Invitrogen™) or other rocker capable of 360° rotation
  2. Magnet for 15 ml tubes
  3. Spectrometer

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Systems Biology > Proteomics > Whole organism

Biochemistry > Protein > Interaction

Molecular Biology > Protein > Protein-protein interaction

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Dear Sir, could you please help us with protocol...
edit 0 Answer 4 Views Mar 24, 2012