AS24 5022 Anti-ABA2 | ABA deficient 2

Confirmed reactivity:	Arabidopsis thaliana
Predicted reactivity:	Brassica napus, Citrus sp., Glycine max, Gossypium sp., Malus domestica, Manihot esculenta, Medicago truncatula, Nicotiana tabacum, Phaseolus vulgaris, Pisum sativum, Populus sp., Theobroma cacao Species of your interest not listed? Contact us
Not reactive in:	No confirmed exceptions from predicted reactivity are currently known

Samples:
1- 10 µg of soluble protein from Arabidopsis thalian Col-0 leaf
2 -10 µg of soluble protein from Arabidopsis thalian aba2-2 leaf
3 - 5 µg of soluble protein from Arabidopsis thalian Col-0 leaf
4 - 5 µg of soluble protein from Arabidopsis thalian aba2-2 leaf
5 - 1 µg purified recombinant ABA2
6 - 500 ng purified recombinant ABA2
7 - 250 ng purified recombinant ABA2
8 - 100 ng purified recombinant ABA2
9 - 50 ng purified recombinant ABA2

5-10 µg/well of frozen total protein extracted from Arabidopsis thaliana leaves (Col-0 & aba2-2). Exact buffer components were: (50 mM Tris-HCl pH 8.0, 10 mM NaCl, 1% SDS, 0.1 mM DTT, 0.5% 2-mercaptoethanol) and denatured with (50 mM Tris-HCl pH 6.8, 6% glycerol, 1.5% w/v DTT, bromophenol blue) at 99 °C/5 min. Samples were separated in the cold on 10 % SDS-PAGE and blotted for 30 min at 20V to PVDF (pore size of 0.2 um), using semi-dry in the cold. Blot was blocked with 5 % milk for: 1h/RT or with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 ON/4°C in TBS-T + 5% skim milk 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 matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 25 000 in TBS-T + 5% skim milk for 1h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent. Exposure time was 5 minutes.

Courtesy of Ben Brookbank from laboratory of prof. Eiji Nambara, University of Toronto, Canada

ABA deficient 2 (ABA2) is involved in Abscisic acid biosynthesis. Catalyzes the conversion of xanthoxin to abscisic aldehyde. Alternative names: Protein abscisic acid deficient 2, Protein glucose insensitive 1, Protein impaired sucrose induction 4, Protein SALOBRENO 3, Protein salt resistant 1, Xanthoxin dehydrogenase

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