LUT1 | Beta-carotene hydroxylase
AS15 3084 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 2000 (WB)|
|Expected | apparent MW||
60.5 | 57 kDa
|Confirmed reactivity||Arabidopsis thaliana|
|Predicted reactivity||Camelia sinensis, Croton stellatopilosus, Daucus carota, Gossypium arboreum, Lycium barbarum, Marchantia polymorpha, Medicago truncatula, Morus notabilis, Oryza sativa, Picea glauca, Ricinus communis, Salvia miltiorrhiza, Selaginella moellendoffoo, rSolanum lycopersicum, Theobroma cacao, Zea mays, Zostera marina|
|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 wilde type (left panel) and lut 1 mutant (right panel), corresponding to 1 µg of chlorophylls, 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: 1,500, 1:3,000, 1:6,000, 1:12,000 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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