Cyt f | Cytochrome f protein (PetA) of thylakoid Cyt b6/f-complex (algal)
AS06 119 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. reinhardtii, C. quitensis Kunt Bartl, C. pumilum, H. vulgare, N. gaditana, N. tabacum, P. patens, S. martensi, Synechocystis 6803 substrain PCC-M, U. prolifera
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GST fusion to cytochrome f from Chlamydomonas reinhartii P23577
34 | 31-32 kDa
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Chlamydomonas reinhardtii samples are lysed cells, Arabidopsis thaliana samples are prepared from thylakoid membranes; Samples were boiled 1 min in 3%SDS,then 10ug of chlorophylls were loaded in each well of a 12% acrylamide gel.
15 µg protein of whole cell extracts precipitated with 80 % acetone followed by re-suspension in 100 mM Tris HCl pH 6.8, 4 % SDS, 20 mM EDTA of Chlamydomonas reinhardtii were loaded on BioRad 4-20% Criterion TGX Precast Mini protein gels. Gels were run at 85V. For electro-blotting wet transfer onto nitrocellulose membrane was performed (105 V for 1h). Overnight blocking with 5% milk in TBS-T in the cold room. Two-hour-long incubation with anti-cyt f antibodies at a dilution of 1:50 000 in TBS-T was followed by washes and incubation with matching anti-rabbit secondary antibodies and development with chemiluminescent detection reagent following manufacture's recommendations. Image was taken using Chemidoc MP Imaging system. Exposure time was in a range of 3 to 10 seconds.
Courtesy of Dr. Angeliki Tsichla, Petroutsos lab, CEA Grenoble, France
Multi-subunit complex of cytb6/f is a crucial component for the photosynthetic electron transport chain of higher plants, green algae and cyanobacteria. This complex is catalyzing oxidation of quinols and the reduction the reduction of plastocyanin. This reaction allows to establish the proton force required for the ATP synthesis. Four major subunits build the complex: the petA gene product corresponding to a c-type cytochrome (cytf), the petB gene product corresponding to a b-type/c’-type cytochrome with three haems (cyt b6), the petD gene product (subunit IV, or suIV), and the petC gene product, corresponding to the Rieske/Iron/sulfur protein.
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