Rhodamine or fluorescein goat antimouse secondary antibodies

For immunodetection of endogenous dystrophin, a small fragment of muscle biopsy was homogenized in extraction buffer (4% SDS, urea 4 M). Lysate protein was loaded on 6% polyacrylamide‐SDS gel and transferred to a nitrocellulose membrane (Schleicher and Schuell). Membranes were probed with the dystrophin Rod domain mouse monoclonal antibody (NCL‐DYS1, dil. 1:200), from Novocastra Laboratories (Newcastle upon Tyne). Actinin (monoclonal antibody, 1:6000 Sigma Aldrich) was used as indicator of protein loaded. The membranes were incubated with rhodamine or fluorescein goat antimouse secondary antibodies (LI‐COR, Lincoln). Immunoreactive bands were visualized by ODYSSEY LI‐COR Model 2800 and quantitated densitometrically using Image J 1.46r software. Dystrophin band was normalized to Actinin band and expressed as percentage with respect to control values.

For immunodetection of endogenous dystrophin, a small fragment of muscle biopsy was homogenized in extraction buffer (4% SDS, urea 4 M). Lysate protein was loaded on 6% polyacrylamide-SDS gel and transferred to a nitrocellulose membrane (Schleicher and Schuell, Keene, Nh, USA). Membranes were probed with the following primary antibodies: dystrophin Rod domain mouse monoclonal antibody (NCL-DYS1, dil. 1:200), dystrophin C-terminus mouse monoclonal antibody (NCL-DYS2, dil. 1:80) all from Novocastra Laboratories (Newcastle upon Tyne, UK).
Actinin (monoclonal antibody, 1:6000 Sigma Aldrich) was used as indicator of protein loaded [14 (link)]. The membranes were incubated with rhodamine or fluorescein goat anti mouse secondary antibodies (LI-COR, Lincoln, NE, USA).
Immunoreactive bands were visualized by ODYSSEY LI-COR Model 2800 and quantitated densitometrically using Image J 1.46r software. Dystrophin bands were normalized to Actinin band and expressed as percentage with respect to control values.