Why do I observe multiple high molecular weight bands on my western blot when using your SOAT1 (E5O4K) Rabbit mAb #84476?

When using overexpressed SOAT1 lysate or a SOAT1 enriched sample (i.e. immunoprecipitation) we observe multiple high molecular weight bands. Specifically for immunoprecipitation experiments, we speculate these bands are due to the experiment being performed in Cell Lysis Buffer (10X) #9803, which contains no SDS or DTT. Therefore, we believe the multiple high molecular weight bands are oligomers of SOAT1 (since they are 90, 135, 180 kDa vs the 45kDa monomer). These oligomers are formed under IP lysate conditions and cannot be broken up by SDS/DTT treatment after formation. In addition, when using normal endogenous cell lines which are lysed in SDS/DTT buffer we do not observe these high molecular weight bands. 

Last updated: September 12, 2024