AS19 4272 Anti-ABI5 | Abscisic acid insensitive 5 (anti-protein antibody)

Confirmed reactivity:	Arabidopsis thaliana
Predicted reactivity:	Brassica oleracea, Solanum lycopersicum, Oryza sativa Species of your interest not listed? Contact us
Not reactive in:	No confirmed exceptions from predicted reactivity are currently known

Samples:
1 - 30 µg total proteins of Arabidopsis thaliana germinating seeds (3-day old); wild type
2 - 30 µg total proteins of Arabidopsis thaliana germinating seeds (3-day old); wild type, treated with 0.5 μM ABA
3 - 30 µg total proteins of Arabidopsis thaliana germinating seeds (3-day old); abi5-8 mutant (a knockdown mutant), treated with 0.5 μM ABA MW Markers: BioRad Precision Plus Protein standards

30 µg (per lane) of total proteins extracted freshly from 3-day-old germinating seeds with the exaction buffer (50 mM Tris-HCl pH 8.0, 200 mM NaCl, 10 mM DTT, 1% (v/v) Triton X-100, Sigma protease inhibitor cocktail) and denatured with 4X SDS sample buffer at 95°C for 5 min. Samples were separated using 10% SDS-PAGE and transferred onto PVDF membrane (0.2 m pore size), using wet transfer. After blocking with 5% milk in PBS-T for 0.5h at RT (room temperature) with agitation, the blot was incubated in the primary antibody at a dilution of 1: 10, 000 (from the initial antibody solution at 1 µg IgG/µl) in PBS-T at 4°C with agitation. The antibody solution was decanted, and the blot was washed 4 times, each for 10 min, in PBS-T at RT with agitation. The blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, 1:20, 000) for 1h at RT with agitation. It was washed and developed with Agrisera ECL SuperBright.

Courtesy of Dr. Hong Wang, University of Saskatchewan, Canada

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.

ABI5 (abscisic acid insensitive 5) is involved in ABA-regulated gene expression during seed development and subsequent vegetative stage and acts as the major mediator of ABA repression of growth. Binds to the embryo specification element and the ABA-responsive element (ABRE) of the Dc3 gene promoter and to the ABRE of the Em1 and Em6 genes promoters.

Alternative names: ABI5, ABA INSENSITIVE 5, GIA1, GROWTH-INSENSITIVITY TO ABA 1, Dc3 promoter-binding factor 1, AtDPBF1, GROWTH-INSENSITIVITY TO ABA 1, bZIP transcription factor 39, AtbZIP39.

1. Stone et al. (2006). KEEP ON GOING, a RING E3 ligase essential for Arabidopsis growth and development, is involved in abscisic acid signaling. Plant Cell. 2006 Dec;18(12):3415-28.

Agrisera Western Blot protocol and video tutorials

Protocols to work with plant and algal protein extracts


Oxygenic photosynthesis poster by prof. Govindjee and Dr. Shevela

Z-scheme of photosynthetic electron transport by prof. Govindjee and Dr. Björn and Dr. Shevela