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RGA | DELLA protein RGA

AS11 1630 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

RGA | DELLA protein RGA in the group Antibodies for Plant/Algal  / Hormones / Biosynthesis/regulation at Agrisera AB (Antibodies for research) (AS11 1630)

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Data sheet Product citations Protocols Customer reviews

Product Information

Immunogen

KLH-conjugated peptide chosen from RGA of Arabidopsis thaliana UniProt: Q9SLH3,TAIR: At2g01570

Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized
Quantity 50 µg
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 : 1000 (WB)
Expected | apparent MW

64 | 64 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Arabidopsis thaliana
Not reactive in

Triticum aestivum

Application examples

Application examples

Application example

western blot using anti-RGA antibodies

40 µg of total protein from 5-d-old dark-grown Arabidopsis thaliana seedlings extracted with 50 mM Tris-HCl pH 7.5,, 10% glycerol, 150 mM NaCl, 0.1% NP-40, 1 mM PMSF, and 1x protease inhibitor cocktail (Roche) were separated on 4-20 % SDS-PAGE and blotted 1h to PVDF. m: mock, P: paclobutyrazol, P+G: PAC+GAs. Blots were blocked with 2% blocking reagent (GE Healthcare) in TBS-T for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT with agitation. 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 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescence detection reagent, according to the manufacturers instructions. Exposure time was 20 seconds in a LAS-3000 Imager (Fuji).

Courtesy of Dr. David Alabadi, IBMCP (CSIC-UPV), Valencia, Spain

westrn blot using anti-RGA antibodies


Arabidopsis thaliana seedlings were ground in liquid nitrogen (100 µl of 2.5 x Laemli for 80-120 mg of homogenized material) and boiled in 2,5x Laemmli Buffer (WITH 60 Mm DTT final concentration) otherwise RGA protein will degrade. Plants were grown on 1/ MS for 15 days and were treated with 1 µM GA for 2 hours (GA+) or without hormone (GA-). Total protein extracts were denatured  for 2 min. at 95°C and were separated on 10% SDS-PAGE and blotted overnight to PVDF using tank transfer. Blots were blocked for 1.5 h with TBS-T containing 5% low fat milk for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h 30min at RT with agitation. The antibody solution was decanted and the blot was washed 5 x 10min 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 1:10 000 for 1h 30 min at RT with agitation. The blot was washed 6 x 10min in TBS-T at RT with agitation and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 5 seconds with West Femto (Pierce). 

Courtesy of Kamila Jaronczyk, IBB, Warsaw, Poland

Additional information

RGA protein is prone to degradation therefore caution has to be taken during protein extraction.

Related products

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AS11 1631 | Anti-GAI | DELLA protein GAI, rabbit antibodies
AS16 3154 | Anti-PIF3 | Phytochrome interacting factor 3, goat antibodies
AS16 3955 | Anti-PIF4 | PHYTOCHROME INTERACTING FACTOR 4, goat antibodies
AS11 1802 | Anti-RGA-like protein | DELLA protein RGL1, rabbit antibodies
AS11 1803
| Anti-RGA-like protein 2 | DELLA protein RGL2, rabbit antibodies

Plant and algal protein extraction buffer

Secondary antibodies

Background

Background DELLA protein RGA is a putative transcriptional regulator that acts as a repressor of the gibberellin (GA) signaling pathway. It is a member of VHIID/DELLA regulatory family and is the most sensitive to GA application, compare to other DELLA proteins.Involved in fruit and flower development.  Synonymes: GAI-related sequence, GRAS family protein 10, AtGRAS-10, Repressor on the ga1-3 mutant, Restoration of growth on ammonia protein 1.

Product citations

Selected references Yan et al. (2020). FKF1 F-box Protein Promotes Flowering in Part by Negatively Regulating DELLA Protein Stability Under Long-Day Photoperiod in Arabidopsis. J Integr Plant Biol . 2020 May 19. doi: 10.1111/jipb.12971.
Ferrero et al. (2019). Class I TCP transcription factors target the gibberellin biosynthesis gene GA20ox1 and the growth promoting genes HBI1 and PRE6 during thermomorphogenic growth in Arabidopsis. Plant Cell Physiol. 2019 Jul 11. pii: pcz137. doi: 10.1093/pcp/pcz137.
Lorrai et al. (2018). Abscisic acid inhibits hypocotyl elongation acting on gibberellins, DELLA proteins and auxin. AoB Plants. 2018 Oct 5;10(5):ply061. doi: 10.1093/aobpla/ply061.
Chahtane et al. (2018). The plant pathogen Pseudomonas aeruginosa triggers a DELLA-dependent seed germination arrest in Arabidopsis. Elife. 2018 Aug 28;7. pii: e37082. doi: 10.7554/eLife.37082.
Shahnejat-Bushehri et al. (2016). Arabidopsis NAC transcription factor JUB1 regulates GA/BR metabolism and signalling. NATURE PLANTS 2: Article number: 16013, 2016.
Crocco et al. (2015). The transcriptional regulator BBX24 impairs DELLA activity to promote shade avoidance in Arabidopsis thaliana. Nat Commun. 2015 Feb 6;6:6202. doi: 10.1038/ncomms7202.
Leone et al. (2014). To grow or defend? Low red : far-red ratios reduce jasmonate sensitivity in Arabidopsis seedlings by promoting DELLA degradation and increasing JAZ10 stability. New Phytol. 2014 Oct;204(2):355-67. doi: 10.1111/nph.12971. Epub 2014 Aug 7.

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