LCY | Lycopene beta-cyclase (chloroplastic)
AS15 3079 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 1000-1 : 128 000 (WB)|
|Expected | apparent MW||
56 | 50 kDa
|Confirmed reactivity||Arabidopsis thaliana|
|Predicted reactivity||Adonis aestivalis var. palaestina, Bixa orellana, Brassica napus, Brassica rapa subsp. pekinensis, Camellia sinensis, Capsicum annuum, Carica papaya, Citrus maxima, Citrus sinensis, Chrysanthemum morifolium, Cucumis sativus, Cucurbita moschata, Daucus carota subsp. sativus, Diospyros kaki, Eriobotrya japonica, Erythranthe lewisii, Gentiana lutea, Glycine soja, Ipomoea sp. Kenyan, Lycium ruthenicum, Medicago truncatula, Morus notabilis, Narcissus pseudonarcissus, Nicotiana tabacum, Populus trichocarpa, Ricinus communis, Rosa rugosa, Salicornia europaea, Sandersonia aurantiaca, Solanum lycopersicum, Taraxacum officinale, Taraxacum officinaleTheobroma cacao, Vitis vinifera|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Selected references||Antibody released in January 2016.
This product can be sold with ProClin if requested
Total protein from Arabidopsis thaliana leaves, wilde type and Lut2 (Arabidopsis thaliana mutant devoid of lycopene-epsilon-cyclase, an enzyme at the same branching point catalyzed by lycopene-beta-cyclase (LCY)) corresponding to 0.5 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 12% 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 from 1:1000 to 1:128000) 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
Mutant devoid of LCY is not available (not viable) and using lut2 mutant allowed to test if the antibody discriminates the two different cyclases.
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