- Home
- Protocols
-
Crosslinking and Immunoprecipitation in Zebrafish
Published: Jan 20, 2012 DOI: 10.21769/BioProtoc.174 Views: 14555
PDF 
Ask a question
Abstract
Immunoprecipitation (IP) is a routine method to detect protein binding and interactions. But the weak binding between two proteins is often hard to detect during regular IP procedure. This protocol offers a crossliking and IP combination method to detect weak binding of proteins in zebrafish embryos.
Background
Materials and Reagents
- NaCl
- KCl
- CaCl2
- Tris
- EDTA
- Glycerol
- Triton X-100
- TBST
- Glycine
- NP-40
- Na deoxycholate
- SDS
- DTSSP (Thermo Fisher Scientific, catalog number: 21578 )
- HEPES (Life Technologies, Invitrogen™, catalog number: 15630 )
- Protease inhibitors (Sigma-Aldrich, catalog number: P2714 )
- Anti-HA affinity matrix (Roche Diagnostics, catalog number: 11815016001 )
- Anti-FLAG M2 affinity gel (Sigma-Aldrich, catalog number: A2220 )
- Modified Ringer’s solution (see Recipes)
- Lysis buffer (see Recipes)
- RIPA (see Recipes)
- Wash buffer (see Recipes)
Equipment
- Kontes tubes
Procedure
Category
Biochemistry > Protein > Immunodetection > Immunoprecipitation
Biochemistry > Protein > Interaction > Crosslinking
Biochemistry > Protein > Interaction > 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
