MnSOD | Manganese superoxide dismutase
AS09 524 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, A. maritima, B.napus, B. oleracea, N. cataria, N. rtanjensis, O. sativa, P. sativum, Salicornia sp., S. tuberosum
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25 | 25 kDa
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5 µg (1,2), 10 µg (3, 4) of total protein from Pisum sativum were separated on 12% SDS-PAGE and blotted 30 min. to PVDF. Blots were blocked (in 5% fat free milk) immediately following transfer in for 1h at RT with agitation. Blots were incubated in the primary antibody at a dilution of 1: 2 000 overnight in 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 4 times for 5 min in TBS-T at RT with agitation. Blots were incubated in secondary antibody (anti- IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:20 000 for 1h at RT with agitation. The blots were washed 4 times for 5 min in TBS-T and 2 times for 5 min in TBS and developed for 1 min with ECL detection reagent according to the manufacturers instructions. Exposure time was 60 seconds.
Courtesy Dr. Elżbieta Romanowska, Warsaw University, Poland
Freshly prepared reducing agent like DTT needs to be used in a sample buffer. Otherwise MnSOD will migrate at 50 kDa.
Plant MnSOD catalyzes the dismutation of superoxide to molecular oxygen and water. One subunit of the homotetramer complex binds 1 Mn cofactor. Antioxidant system works as a defense against oxidative stress. SOD (superoxide dismutase) catalyzes the dismutation of superoxide into oxygen and H,O,. SODs are classified, according to their metal cofactor, as FeSOD, MnSOD, or Cu / ZnSOD. Chloroplasts generally contain Cu/ZnSOD and, in a number of plant species, FeSOD. Synonymes: MSD1, SODA, SOD3
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