Thermo Fisher Scientific
Dynabeads Protein A for Immunoprecipitation
10002D
Info: Supplier Page
The WNT pathway is essential for embryonic development and adult tissue homeostasis. Deregulation of this pathway results in cancers of the stomach, intestine, breast, etc. I am interested in studying the molecular mechanism of oncogenic WNT signal transduction to identify potential targets for therapeutic intervention of colon cancer. The goal of this experiment is to immunoprecipitate endogenous AXIN1 using an antibody and Dynabeads Protein A.
Immunoprecipitation
RKO whole cell extract
Overnight incubation with AXIN1 antibody-Protein A bead complex
Whole cell extracts from RKO cells were prepared and 3 mg of extract was incubated with 2 µg of AXIN1 antibody-10 µL Dynabeads Protein A bead complex overnight at 4 deg C. Beads were washed five times with lysis buffer and immune complexes were eluted in 2x SDS sample buffer (95 deg C heating for 5 min). Proteins were resolved on a 4-12% Bis-Tris gel and transferred to a nitrocellulose membrane.
N/A
Washing steps are relatively easy with magnetic beads.
None.
Excellent reagent for antibody capture in immunoprecipitation application.