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After six times washing prior to incubation for 5 min with chemiluminescent peroxidase substrate, the blot was visualized with a Diana III cooled CCD-camera imaging system (Raytest, Straubenhardt, Germany).
The blot was visualized with chemiluminescence (ECL, GE Healthcare Life Sciences, Buckinghamshire, UK).
The membrane was washed extensively in PBST buffer to remove any excess secondary antibodies, and the blot was visualized with enhanced chemiluminescence reagent (GE Healthcare, Piscataway, NJ).
The filter was washed extensively in PBST buffer to remove the secondary antibody, and the blot was visualized with ECL reagent.
Fluorescent secondary antibodies (anti-mouse IRDye 800 and anti-rabbit IRDye 680, LI-COR Biosciences) were added and the blot was visualized with the Odyssey Infrared Imaging System (LI-COR Biosciences).
Finally, blots were visualized with Western Blotting Luminol Reagent (Santa Cruz Biotechnology) and densitometry was performed with ImageJ software (http://imagej.nih.gov/ij/).
Blots were visualized with enhanced chemiluminescence (ECL™ Plus Western Blotting Detection Kit; GE Healthcare, Piscataway, NJ).
After the incubation with horseradish peroxidase-conjugated secondary antibody (Santa Cruz Biotech), the blots were visualized with enhanced chemiluminescence sc-20488, Santa Cruz Biotech).
Gels were transferred to nitrocellulose membrane (Invitrogen), and blots were visualized with the ECL chemiluminescent detection system (GE Healthcare, Piscataway, NJ).
For caldesmon (CaD) and rat SmAV protein content quantification, the blots were visualized with a SuperSignal West Pico peroxide solution (Pierce, Rockford, IL).
After incubation in horseradish peroxidase secondary antibody, blots were visualized with ECL substrate (Amersham Bioscences, NJ) and films were quantified by densitometry with an image analyzer system (Syngene,UK).
