- Home
- Protocols
-
Rabbit IgG Conjugation to Dynabeads
Published: May 5, 2011 DOI: 10.21769/BioProtoc.63 Views: 17020
PDF 
Ask a question
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
Immunoglobulin G
Conjugation
Immunoprecipitation
Affinity purification
Background
Materials and Reagents
- Invitrogen Conjulation kit (143.11D), contains C1, C2, HB, LB, SB solutions
- IgG (Sigma-Aldrich, catalog number: I5006-10 mg )
- Dynabeads M-270 Epoxy 300 mg (Life Technologies, Invitrogen™)
- Phosphate buffered saline (PBS) tablet
- 30% NaN3 (fresh)
- 20% Tween 20
Equipment
- Hula mixer (Life Technologies, Invitrogen™) or other rocker capable of 360° rotation
- Magnet for 15 ml tubes
- Spectrometer
Procedure
Category
Systems Biology > Proteomics > Whole organism
Biochemistry > Protein > Interaction > Protein-ligand interaction
Molecular Biology > Protein > Protein-protein interaction
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this article to respond.
Post a Question
