GID2 | F-BOX protein GID2 (SLEEPY 1)
AS13 2638 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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17.4 | 18 kDa (Arabidopsis)
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15µg of total protein from inflorescence of Col0, spy mutant, and 35S-SPY extracted with extraction buffer (DTT 100uM, Tris pH 6,8 67,5mM, Urea 4M, SDS 3%,glycerol 3% and bromophenol 0,1%) were separated on 10% SDS-PAGE and blotted 1h to PVDF. Blots were blocked with milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5000 overnight at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody diluted to 1:10 000 in TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 2 min with chemiluminescent detection reagent, according to the manufacturer's instructions. Exposure time was 30 seconds.
Courtesy Dr. Patrick Achard, CNRS, France
GID2 | F-BOX protein GID2 (SLEEPY 1) is an essential component of the SCF-type E3 ligase complex, which positively regulates the gibberellin signaling pathway. It mediates the ubiquitination and subsequent degradation of DELLA proteins (GAI, RGA and RGL2). Localized in nucleus and expressed in all tissues tested so far.