Anti-ALDH2 | Aldehyde dehydrogenase 2 (plant)

Arachis hypogaea, Brachypodium distachyon, Brassica napus, Capsicum annuum, Citrus sp., Crocus sativus, Cucumis sativus, Glycine max, Hordeum vulgare, Malus domestica, Medicago truncatula, Nicotiana tabacum, Oryza sativa, Phaseolus vulgaris, Pisum sativum, Populus sp., Saccharum sp., Solanum lycopersicum, Solanum tuberosum, Theobroma cacao, Sorghum bicolor, Triticum sp., Vitis vinifera, Zea mays

10.8 µg/well of total protein extracted from Nicotiana tabcum mitochondria. Exact buffer components were: 100 mM Tris-HCl pH 7,4; 2 mM EDTA; 300 mM Sucrose; 0,6% PVPP; 1 mM PMSF; 1% Protease Inhibitor Cocktail and denatured with exact buffer components at 98°C/5 min.  Samples were separated in the room temperature on 4-15 % SDS-PAGE and blotted for 1 h ? PVDF or nitrocellulose (pore size of 0.22um), using: semi-dry transfer. Blot was blocked with 3% BSA for 4°C/ON with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2500 for 1h/RT with agitation in TBS-T. 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 for 1h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagent. Exposure time was 1 minute.

Note: protein load/well can be decreased to 2 µg/well and the antibody can be used at higher dilution. 

Courtesy Yufeng Guan, PhD Candidate, Department of Plant Ecophysiology, Faculty of Biology, UAM, Poznań, Poland