LUT5 | beta-carotene hydroxylase
AS15 3085 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 2000-1 : 8000 (WB)|
|Expected | apparent MW||
66.8 | 64 kDa
|Confirmed reactivity||Arabidopsis thaliana|
|Predicted reactivity||Arabidopsis thaliana|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
to be added when available, antibody released in January 2016.
Total proteins from Arabidopsis thaliana leaves, corresponding to 1 µg of chlorophylls of wilde-type (wt) and Lut5 mutant, were extracted with loading buffer (10% glycerol, 62.5 mM Tris pH 6.8, 2% SDS, 5% β-mercaptoethanol) and denatured at 100°C (boiling water) for 1 min. Proteins were separated on 15% SDS-PAGE (Laemly) and blotted 1h to PVDF using tank transfer. Blots were blocked with blocking solution (PBS 1X, 0.2% w/v Tween, 5% powder milk) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody diluted in blocking solution, at a dilution of 1: 2,000, 1:4,000, 1:8,000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in blocking solution at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG alkaline phosphatase conjugated) diluted to 1:30 000 in blocking buffer for 1h at RT with agitation. The blot was washed 2 times for 10 min in blocking solution and once with PBS 1X solution for 10 min, then developed in developing buffer NBT/BCIP by manual agitation.
Courtesy of Stefano Cazzaniga, University of Verona, Italy
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