FNR | Ferredoxin-NADP+-oxidoreductase

 10 ng of recombinant Chlamydomonas reinhardtii FNR (1), thylakoid preparation (1 µg chl) from Chlamydomonas reinhardtii cc 124 (2), thylakoid preparation (1 µg chl) of Chlorella sorokiniana (3), thylakoid preparation (1 µg chl) of Synechococcus PCC 6803 (4) incubated with urea based sample buffer over night at 25°C were separated on Bolt 4-12 % Bis-Tris Plus gels (Novex, Life Tech) and blotted 1h to nitrocellulose using iBlot Gel Transfer Stacks Nitrocellulose, Mini (Novex, LifeTech). Blots were blocked with iBind (LifeTech) with iBind solution kit (Novex, LifeTech). Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT with agitation. There is no washing steps using this set up. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602 ) diluted to 1:40 000 in for 1h at RT with agitation. The blot was washed as above and developed for with chemiluminescent detection reagent, according to manufacture recommendations. Exposure time was 10 min.

Courtesy of M.Sc. Pini Marcu, Tel Aviv University, Israel

Proteins were isolated from wt Arabidopsis thaliana and fnr1 plants with 3X LB (6 M urea, 12% SDS, 30% glycerol, 100 mM DTT, 150 mM Tris pH7.0, 0.8% Comassie G-250). 10 µg of total proteins from leaves or roots were loaded into each lane and separated on 12% SDS-PAGE, and then blotted overnight onto PVDF membrane. Membranes were blocked with milk powder for 2 h and then incubated in the primary antibody solution overnight, which was then decanted and the membrane was washed 3 times for 5 min in TBST. Membrane was incubated at RT for 1 hour in 1:10 000 goat anti-Rabbit secondary antibody from Agrisera, followed by washing steps as above. Membrane was developed for 2 min with highest sensitivity chemiluminescent detection reagent according to the manufacturer’s instructions and recorded using FujiFilm CCD camera with 10 s increment time for around 190 s.

Arabidopsis has four FNR proteins, two of them are found in leaves (LFNR1 and LFNR2) while the other two in roots (RFNR1 and RFNR2). Absence of one of leaf FNR results in a decrease in the amount of FNR while absence of both of them is lethal.

For detection in Chlorella sorokiniana, Synechococcus PCC 6803 higher load per well needs to be applied.

This antibody recognizes all Arabidopsis thaliana FNR isoforms.

This product can be sold with ProClin if requested

FNR (ferredoxin-NADP+-oxidoreductase) catalyzes reduction of NADP+ using Fd that has accepted electrons from photosystem II in the final step of linear photosynthetic electron transport. In higher plants FNRs are encoded by a small multiple gene family (two chloroplast-targeted FNR in Arabidopsis and rice and three isoenzymes in maize). All forms are evenly distributed between the thylakoids and stroma.