DRB4 | Double-stranded RNA-binding protein 4
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|Expected | apparent MW||
|Predicted reactivity||Brassica napus, Brassica oleracea, Camelina sativa, Capsella rubella, Eutrema salsugineum|
|Not reactive in||
No confirmed exceptions from predicted reactivity are currently known.
To be added when available.
To be added when available, antibody released in October 2016.
100 µg of total protein from Arabidopsis wt, drb4, and 35S-DRB4 leaves extracted with isolation buffer (50 mM Tris-HCl, pH 7.5, 10% glycerol, 150 mM NaCl, 10 mM MgCl2, 5 mM EDTA, 5 mM DTT, and 1 X protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO) and denatured by boiling at 100°C. The proteins were separated on 10% SDS-PAGE and blotted 1.5 hrs to PVDF membrane using wet transfer. Blots were blocked with 5% milk powder for 30 min at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1,000 for 60 min at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in phosphate buffer (containing 0.1% Tween 20) at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG alkaline phosphatase conjugated) diluted to 1:10,000 for 120 min at RT with agitation. The blot was washed as above and developed with NBT/BCIP (Fisher Chemicals) using Alkaline phosphatase detection system. The DRB4 specific band was seen within 20 mins and showed a better signal when allowed to develop for longer duration (overnight in this case).
Courtesy of Dr. Gah-hyun Lim and Prof. Pradeep Kachroo, University of Kentucky, Lexington, USA
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