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These antibody protein complexes on the blots were detected using ECL-plus Western blotting detection reagents (GE Healthcare) following incubation with anti-mouse IgG horseradish peroxidase (GE Healthcare) for 60 min at room temperature.
Bound antigens on western and dot blots were detected using the ECL Advance chemiluminescence detection kit (GE Healthcare, Piscataway, NJ, USA).
A secondary antibody, donkey anti-rabbit HRP-linked IgG, 1 2000 (Amersham Biosciences, Buckinghamshire, UK), was added for one hour and blots were detected using an enhanced chemiluminescence detection kit (Amersham Biosciences, Buckinghamshire, UK).
A secondary antibody, donkey anti-rabbit horseradish peroxidase-linked IgG (1 2,000; Amersham Biosciences), was added for 1 hour and blots were detected using an enhanced chemiluminescence detection kit (Amersham Biosciences).
Blots were detected using an enhanced chemiluminescence (ECL) detection kit (Amersham).
Western blots were detected using Clarity Western ECL substrate (Bio-Rad) and imaged with the ChemiDoc Touch Imaging System (Bio-Rad).
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After a final wash with TBST, the immune-reactivity of the blots was detected using an enhanced chemiluminescence detection system (Amersham, Piscataway, NJ).
After a final set of washes, the immunoreactivity of the blots was detected using an enhanced chemiluminescence detection system (Amersham, Buckinghamshire, UK).
After washing, the immunoreactivity of the blots was detected using an ECL Plus Western Blotting Detection System (GE Healthcare, Tokyo, Japan).
The presence of citrullinated proteins on the nitrocellulose blots was detected using the anti-modified citrulline (AMC) detection kit (Upstate, Charlottesville, VA, USA) according to the manufacturer's protocol.
Antibody-bound proteins on the blot were detected using enhanced chemiluminescence on X-ray film.
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