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LUT5 | beta-carotene hydroxylase

AS15 3085 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

LUT5 | beta-carotene hydroxylase  in the group Antibodies for Plant/Algal  / Photosynthesis  / Carotenoid metabolism at Agrisera AB (Antibodies for research) (AS15 3085)

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Product Information

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 tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications Western blot (WB)
Recommended dilution 1 : 2000-1 : 8000 (WB)
Expected | apparent MW

66.8 | 64 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Arabidopsis thaliana
Not reactive in diatoms

Application examples

Application examples application example

western blot using anti-LUT5 antibodies

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

Additional information

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Background

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.

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