AS12 1871 Anti-ABI2 | Abscisic acid insensitive 2

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

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.

To detect endogenous ABI2 plant material needs to be subjected to stress before harvesting.

Protein Abscisic acid-insensitive 2 (ABI2) is a represor of the ABA signaling pathway, regulating various ABA responses like: stomatal closure, osmotic water permability of the plasma membrane, high light stress, response to glucose, seed germination and inhibition of vegetative growth. Involved in acquired thermotolerance. Alternative names: ABA INSENSITIVE 2, ABI2, ATABI2, Protein phosphatase 2C 77, AtPP2C77, At5g57050, MHM17_19, MHM17.19, PP2C ABI2.

1. 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.