Anti-LUT5 | beta-carotene hydroxylase
AS15 3085 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
- Product Info
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Immunogen: His-tagged, recombinant LUT5 of Arabidopsis thaliana, overexpressed in E.coli, UniProt: Q93VK5,TAIR: AT1G31800
Host: Rabbit Clonality: Polyclonal Purity: Serum Format: Lyophilized Quantity: 50 µl Reconstitution: For reconstitution add 50 µl of sterile water Storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube. Tested applications: Western blot (WB) Recommended dilution: 1 : 2000-1 : 8000 (WB) Expected | apparent MW: 66.8 | 64 kDa - Reactivity
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Confirmed reactivity: Arabidopsis thaliana Predicted reactivity: Arabidopsis thaliana Not reactive in: Diatoms - Application Examples
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Application example
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 - Background
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Background: LUT5 (beta-carotene hydroxylase) is a heme-containing cytochrome P450 involved in the biosynthesis of xanthophylls. Enzyme is specific for beta-ring hydroxylation of alpha- and beta-carotene and displays a low activity toward the epsilon-rings of alpha-carotene.
Alternative names: Cytochrome P450 97A3, Protein LUTEIN DEFICIENT 5, chloroplastic. - Protocols
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Accessories
AS09 602 | Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)
AS15 3091 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Brachypodium distachyon, Deschampsia antarctica, Oryza sativa
AS15 3084 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana