DET1 | Regulator of the proteasomal degradation of LHY

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AS15 3082 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana


26 st
Item No:
AS15 3082

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product information
Background DET1 (regulator of the proteasomal degradation of LHY) is a component of light signal transduction (repression of photomorphogenesis in darkness) machinery, localized in nuclei. Alternative name: Protein DEETIOLATED 1.

Recombinant, full length DET1 of Arabidopsis thaliana, overexpressed in E.coli, UniProt: P48732, TAIR: AT4G10180

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl 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)
Related products

AS13 2659 | anti-CCA1 | Circadian clock assiociated 1, rabbit antibodies

AS13 2661 | anti-LHY | Late elongated hypocotyl, rabbit antibodies

collection of antibodies to proteins involved in signal transduction

Plant protein extraction buffer

Secondary antibodies

Additional information
application information
Recommended dilution 1 : 2500 (WB)
Expected | apparent MW

62 | 62 kDa

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Capsicum annuum, Glycine max, Medicago truncatula, Oryza sativa, Populus trichocarpa, Ricinus communis, Solanum lycopersicum, Solanum tuberosum, Theobroma cacao, Zea mays, Zostera marina
Not reactive in No confirmed exceptions from predicted reactivity are currently known.
Additional information
Selected references Castells et al (2011). The conserved factor DE-ETIOLATED 1 cooperates with CUL4-DDB1DDB2 to maintain genome integrity upon UV stress. EMBO J. 2011 Mar 16;30(6):1162-72. doi: 10.1038/emboj.2011.20. Epub 2011 Feb 8.

application example

western blot using anti-DET1 antibodies

Recombinant protein (0.5-1-1.5-2 µg of protein) and total proteins from Arabidopsis thaliana whole leaves from plants grown in control light (C), low light (LL) and high light (HL), 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  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 1: 25,000 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 developing buffer NBT/BCIP by manual agitation. 

Courtesy of Stefano Cazzaniga, University of Verona, Italy

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