GAI | DELLA protein GAI
AS11 1631 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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20 µg of total protein from Arabidopsis thaliana Columbia-0 plants grown on agar plates (MS 1 % sucrose) in LD conditions (16 h light, 8 h dark). Protein extraction was done using TRIZOL protocol on samples taken every 4 hours from the time point lights were switched on (Zeitgeber Time 0). Marker was 10 µl of BLUEstain™ Protein ladder, 11-254 kDa (GoldBio). Samples were separated on 12 % SDS-PAGE run at 200 V on Mini-Protean and blotted 1h to nitrocellulose. Blots were blocked with TBST (TBS+0.1 % Tween, 5 % non-fat, dry milk) for over night at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 2h at RT with agitation in TBS-T. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:00 000 in for 1h at RT with agitation. The blot was washed as above and developed with BIO-RAF Chemi-doc. Exposure time was 5 minutes.
Courtesy of Dr Federico Valverde, CSIC – University of Seville, Spain
Protocol for protein extraction from seeds can be requested here.
GAI is a probable transcriptional regulator that acts as a repressor of the gibberellin (GA) signaling pathway. Less sensitive to GA. Synonymes:GRAS family protein 3, AtGRAS-3, Gibberelic acid-insensitive mutant protein, Restoration of growth on ammonia protein 2.
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