Thank you for your question. We recommend blocking with Blotto (5% nonfat milk in TBST) for one hour at room temperature, but you can try overnight at 4C if you are experiencing high background. We then recommend diluting the primary antibody in fresh Blotto at a dilution of 1:200 and incubating for one hour at room temperature. We load 50 µg sample protein. And to help reduce non-specific banding, we recommend using our specific mouse IgG kappa binding protein (sc-516102) in place of a conventional anti-mouse IgG secondary antibody. A link to our complete recommended protocol is provided below.
If you continue to have trouble, please contact our Technical Service Department and we can assist you with troubleshooting the antibody.
Date published: 2021-01-05