FNR3 | Ferredoxin NADP Reductase, isoprotein 3 (leaf)
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2 μg/well of leaf total protein of Arabidopsis thaliana wild type leaf (1), Zea mays leaf (2) were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE. For IP, 150mM NaCL, 1% Triton X-100, 50 mM Tris-HCl (pH 8.0) and denatured with 4X SDS buffer at 95°C for 5 min. Samples were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 in TBS-T for 1-2h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation.
Molecular weight of mature forms of maize L-FNRs:
34.97 kDa (FNR1, Zea mays), 35.57 kDa (FNR2, Zea mays), 34.7 kDa (FNR3, Zea mays)
Okutani et al. (2005). Three Maize Leaf ferredoxin:NADPH Oxidoreductases Vary in Subchloroplast Location, Expression, and Interaction With Ferredoxin. Plant Physiol. 2005 Nov;139(3):1451-9. doi: 10.1104/pp.105.070813.
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