Why do I see a doublet when using your Na,K-ATPase Antibody #3010?

We have found that a doublet may appear if samples are boiled prior to gel loading when performing a western blot with our Na,K-ATPase Antibody #3010.  The protein structure of this target is extremely sensitive to denaturing elements such as temperature, pH, ionic strength, presence of native lipids, cholesterol levels, and detergent concentration. We believe that the upper band is caused by protein aggregation due to this sensitivity and the location of the #3010 antigen in the transmembrane region of the protein. 
Therefore, we recommend omitting the boiling step during lysate preparation when using this antibody, which should eliminate the background band that sometimes appears above the target. 

Last updated: March 6, 2023