Phly | DNA photolyase (At4g25290) (N-terminal part)
AS15 2863 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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1 : 1000 with ECL (WB)
|Expected | apparent MW||
78 | 90 kDa
|Not reactive in||
no confirmed exceptions from predicted reactivity are currently known
to be added when available antibody available in March 2016.
2,5 µg of total protein from Arabidopsis thaliana wilde type darkness (1), wilde type light (2) and insertion mutants: SALK_056328C darkness (3), SALK_056328C light (4), extracted with 0.1 M Tris-HCl pH 8.5, 4% SDS, 2% (v/v) 2-mercaptoethanol, 2 mM phenylmethylsulfonyl fluoride and denatured with Laemmli buffer at 95oC for 10 min were separated on 12% SDS-PAGE and blotted 2h to PVDF using semi-dry transfer. Blots were blocked with 5% milk PBS-T (Tween 0.5%) for 30 min. at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 overnight at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in 5% milk PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera) diluted to 1:25 000 in 5 %milk PBS-T for 1h at RT with agitation. The blot was rinsed briefly twice, then washed 3 times for 10 min in PBS-T at RT with agitation. Blot was developed for 5 min with Clarity™ Western ECL Blotting S ubstrate (Biorad). Exposure time was 5 minutes.
Courtesy of Dr. Justyna Łabuz, Department of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Poland
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