GAI | DELLA protein GAI
AS11 1631 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Product Information
Immunogen
KLH-conugated synthetic peptide, chosen from Arabidopsis thaliana GAI protein sequence UniProt: Q9LQT8, TAIR: At1g14920
Host
Rabbit
Clonality
Polyclonal
Purity
Affinity purified serum in PBS, pH 7.4
Format
Lyophilized
Quantity
50 µg
Reconstitution
For reconstitution add 25 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 : 1000 (WB) on transfected Arabidopsis thaliana protoplasts
Expected | apparent MW
58.9 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana
Not reactive in
Solanum lycopersicum
Application examples
Application examples
Application example
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
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
Additional information
Homogenization with thiourea and bead beater.
Protocol for protein extraction from seeds can be requested here.
Protocol for protein extraction from seeds can be requested here.
GAI protein is present in very low levels therefore specific material should be used for analysis as well as chemiluminescence detection reagent in extreme low femtogram range, as AgriseraECLSuperBright.
Background
Background
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.
Product citations
Selected references
Shahnejat-Bushehri et al. (2016). Arabidopsis NAC transcription factor JUB1 regulates GA/BR metabolism and signalling. NATURE PLANTS 2: Article number: 16013, 2016.
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