GAI | DELLA protein GAI

AS11 1631 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

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

Additional information

Homogenization with thiourea and bead beater.

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.

Related products

Plant and algal protein extraction buffer

Secondary antibodies

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.