NdhB | NAD(P)H-quinone oxidoreductase subunit 2 (chloroplastic)
AS16 4064 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Zea mays
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5µg of total protein from Zea mays leaf seedling tissue was extracted with protein extraction buffer (100 mM Tris pH7.5, 10% glycerol, 1 mM EDTA, 1mM EGTA, 2mM pMSF, 0.002 mg leupeptin, 0.002 mg pepstatin A, 2.5 mM DTT) and denatured in 1XPSB (33 mM Tris pH 6.8, 50 mM 2-Mercaptoethanol, 2% SDS, 10% glycerol, 0.1% bromophenol blue) at 70C for 5 min, separated on 4-20% Tris-Glycine SDS-PAGE and blotted overnight to nitrocellulose using tank transfer. Blots were blocked with 10% milk in 1XTBSt for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for 1h at RT with agitation in 10% milk in 1XTBS-T. 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 1:10,000 in 10% milk in 1XTBS-T for 1h at RT with agitation. The blot was washed as above and developed for 2 min with chemiluminescent detection reagent.
Courtesy Rosalind Carrier, University of Oregon, USA
NAD(P)H-quinone oxidoreductase subunit 2 is involved in the transfer of electrons from NAD(P)H:plastoquinone, via FMN and iron-sulfur centers, to quinones in the photosynthetic chain and possibly in a chloroplast respiratory chain. Alternative names: NAD(P)H dehydrogenase, subunit 2, NADH-plastoquinone oxidoreductase subunit 2.