DET1 | Regulator of the proteasomal degradation of LHY
AS15 3082 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Product Information
Immunogen
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 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 : 2500 (WB)
Expected | apparent MW
62 | 62 kDa
Reactivity
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
Species of your interest not listed? Contact us
Species of your interest not listed? Contact us
Not reactive in
No confirmed exceptions from predicted reactivity are currently known.
Application examples
Application examples
application example
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
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
Additional information
Additional information
This product can be sold containing ProClin if requested.
Background
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.
Alternative name: Protein DEETIOLATED 1.
Product citations
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.
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