Home >
Community >
Unknown bands (at 150 kDA) during western blotting of beta catenin?
Upvote
20
Downvote
+ Western blot
+ Antibodies
Posted by
Laurie Keller
Unknown bands (at 150 kDA) during western blotting of beta catenin?
Dear all,some time ago I did some western blots to determine the beta catenin (active) content in various samples (primary, human VSMC & Saos-2 cells) with different treatments. Strangely, an unknown band (150 kDa) always appears above the expected band of 92 kDA for active beta-catenin. I already was in contact with the customer service that supplied my b-cat antibody (05-665; Merck) and they also confirmed this issue. However, I think it cant be a dimer (would expect that at 200 kDa, not 150 kDa) or phosphorylated b-catenin. Does anyone maybe has an idea what it could be? Due to those circumstances, I am also not sure if I still can use those blots to quantifiy for beta-catenin?Just some details: I am using a Nitrocellulose membrane, goat anti-mouse hrp as sec. antibody, milk blocking buffer, and the underlying systems are from BioRad. For the measurement, I am using the Femto/Pico substrates from Thermo Fisher.Thank you very much for your help in advance!Best, Stefan
