ABI2 | Abscisic acid insensitive 2
AS12 1871 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Additional information||ABI2 antibodies recognize recombiant StrepTag-ABI2, ABI2-GST, His-ABI2.
ABI2 protein is easily degraded therefore extraction buffer needs to contain protease inhibotors, example of such inhibitor coctail can be found here.
|Selected references||Mitula et al. (2015). Arabidopsis ABA-Activated Kinase MAPKKK18 is Regulated by Protein Phosphatase 2C ABI1 and the Ubiquitin-Proteasome Pathway. Plant Cell Physiol. 2015 Dec;56(12):2351-67. doi: 10.1093/pcp/pcv146. Epub 2015 Oct 6.|
GST-ABI1 and GST-ABI2 were purified on glutathione sepharose and separated on 10% SDS-PAGE and blotted 1h to PVDF (semi-dry). Blots were blocked with 3% semi-skimmed milk for 30 min. at room temperature (RT) with agitation. Blots were incubated with the anti-ABI2 primary antibody diluted to 1: 1000 for 30 minutes at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 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 1:50 000 for 30 min. at RT with agitation. The blot was washed as above and developed for 1 min with ECL according to the manufacturer's instructions. Exposure time was 5 min.
Multiple bands are a result of degradation of GST-ABI2 protein.
Courtesy of Małgorzata Tajdel, from Dr. Agnieszka Ludwików labolatory, Adam Mickiewicz University, Poland
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