DISCONTINUED Cyt f | Cytochrome f protein (PetA) of thylakoid Cyt b6/f-complex (higher plants)

AS08 306 | Clonality: Polyclonal | Host: Rabbit | reactiviy: A. thaliana, E. crus-galli, H. vulgare, N. tabacum, P. miliaceum, P. abies, P. sativum, T. elongatus, S. lycopersicum, Synechocystis sp. PCC6803, Z. mays

This antibody is replaced by product AS20 4377

DISCONTINUED Cyt f | Cytochrome f protein (PetA) of thylakoid Cyt b6/f-complex (higher plants) in the group Antibodies Plant/Algal  / Photosynthesis  / Electron transfer at Agrisera AB (Antibodies for research) (AS08 306)

Data sheet Protocols Add review
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Reactant: Bienertia sinuspersici

Application: Western Blotting

Pudmed ID: 22394490

Journal: Plant Methods

Figure Number: 8B

Published Date: 2012-03-06

First Author: Lung, S. C., Yanagisawa, M., et al.

Impact Factor: 5.139

Open Publication

Silver staining and Western blot analyses of protein extracts from the isolated dimorphic chloroplasts. Two micrograms of proteins from total chloroplasts (lane 1), peripheral chloroplasts (lane 2) and central chloroplasts (lane 3) were separated on SDS/12% PAGE. Molecular weights are shown on the left in kilodaltons. (A) Total proteins were visualized by silver staining. Polypeptides of the highly abundant pyruvate orthophosphate dikinase (PPDK), Rubisco large-subunit (LSU) and small-subunit (SSU) are indicated. Other bands of significantly different intensities between the dimorphic chloroplasts are indicated by arrowheads. (B) Resolved proteins were electroblotted onto nitrocellulose membranes and probed with antibodies against PPDK, LSU, Photosystem II manganese-stabilizing protein (PsbO), cytochrome f (Cyt-f), or Photosystem I subunit II (PsaD). (C) The immunoblots were analyzed by densitometric quantification. The abundance of each protein in peripheral (P-Chls) and central (C-Chls) chloroplasts was calculated relative to that of total chloroplasts and shown with standard errors, after three (PsbO) or four (others) independent experiments.

Reactant: Mus musculus (House mouse)

Application: Western Blotting

Pudmed ID: 31245706

Journal: Plant Direct

Figure Number: 5A

Published Date: 2018-02-01

First Author: Rantala, S. & Tikkanen, M.

Impact Factor: None

Open Publication

Stepwise detachment of photosynthetic protein complexes from the thylakoid membrane of WT, stn7, stn8, stn7stn8, and tap38/pph1. Thylakoid membranes of WT, stn7, stn8, stn7stn8, and tap38/pph1 plants harvested from moderate growth light (GL, 120 ?mol photons m?2 s?1) or after high light illumination (HL, 600 ?mol photons m?2 s?1 for 2 hr) were isolated and solubilized with 0%, 0.25%, 0.5%, 1%, and 2% DIG. Equal volumes of the soluble supernatant (S) and insoluble pellet (P) fractions were loaded and separated in SDS?PAGE followed by immunodetection of (a) proteins D1, PSAB, CYTF, and ATPF representing the protein complexes PSII, PSI, LHCII, Cyt b6f, and ATP synthase, respectively, as well as (b) proteins LHCB1, P?LHCB1, LHCB2, P?LHCB2, and LHCB3, representing the different subunits of the LHCII complexes. The differences in mutants with respect to WT as well as the differences in WT in response to Hl are marked with white boxes. Representative data from three different biological replicates are shown

Reactant: Solanum lycopersicum (Tomato)

Application: Western Blotting

Pudmed ID: 34439953

Journal: Biology (Basel)

Figure Number: 9A

Published Date: 2021-07-28

First Author: Trojak, M. & Skowron, E.

Impact Factor: None

Open Publication

Western Blot analyses (a) and densitometric analyses of PsbS (b), VDE (c), PGRL1 (d), and cytf (e) proteins in leaves of tomato plants (Solanum lycopersicum L. cv. Malinowy Ozarowski) grown under different light conditions (see Material and Methods for details). The bands were normalized to the appropriate ? subunit of ATP synthase (ATPB) band (loading control, Figure S4) (a). The bar diagrams (b–e) represent pixel volumes (densitometric analyses) of proteins in samples. Each value represents the mean ą SD (n = 3) considering the control sample (WL) value as 1 (100%). Different letters indicate significant differences between treatments (p = 0.05) with a Tukey’s HSD test. MW—molecular weight.

Picture (footer): Application example

Western blot using anti-PetA antibodies

1.0 µg of chlorophyll from pea (C3 plant) and from mesophyll (M) and bundle sheath (BS) thylakoids of various treatments of Zea mays, Echinochloa crus-galli, Panicum miliaceum (C4 plants) extracted with 0.4 M sorbitol, 50 mM Hepes NaOH, pH 7.8, 10 mM NaCl, 5 mM MgCl2 and 2 mM EDTA. Samples were denatured with Laemmli buffer at 75 0C for 5 min and were separated on 12% SDS-PAGE and blotted 30 min to PVDF using wet transfer. Blot was blocked with 5% milk in TBS for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 3000 overnight at 40C with agitation in 1% milk in TBS-T. The antibody solution was decanted and the blot was washed 4 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, Lot 1711) diluted to 1:25 000 in 1 % milk in TBS-T for 1h at RT with agitation. The blot was washed 5 times for 5 min in TBS-T and 2 times for 5 min in TBS, and developed for 1 min with 1.25 mM luminol, 0.198 mM coumaric acid and 0.009% H2O2 in 0.1 M Tris- HCl, pH 8.5. Exposure time in ChemiDoc System was 125 seconds.

Courtesy of Dr. Wioleta Wasilewska-Dębowska, Warsaw University, Poland

Western blot using anti-PetA antibodies

Thylakoid membranes (10 µg of total chlorophyll) extracted freshly from Hordeum vulgare leaves with 100 mM HEPES-KOH (pH 7.5), 0.3 M sorbitol, 2 mM EDTA, and 1mM MgCl2 and denatured with a Laemmli buffer at 80°C for 5 min were separated on 12% SDS-PAGE and blotted 1 h to nitrocellulose (pore size of 0.2 um), using semi-dry transfer. Blot was blocked with 4% milk for 2 h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:3000 (PC and PetA, simultaneous western blot detection for both antibodies at the same time) for 1 h/RT with agitation in PBS-T. The antibody solution was decanted and the blot was rinsed briefly, then washed 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25000 in for 1 h/RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent according to manufactures recommendations. Exposure time was 30 seconds. Simultaneous western blot detection can be applied if MW of detected proteins differs in min. 20 kDa.

Courtesy Dr. Anja Liszkay, CNRS, France
calculated | apparent molecular mass [kDa]:

31-32 kDa

Clonality: Polyclonal
Format: Lyophilized
Host: Rabbit

maize cytochrome f purified from chloroplasts, including a final gel purification on a denaturing gel. protein used to elicit this antibody is conserved in Arabidopsis thaliana cyt f UniProt: P56771, TAIR: AtCg00540

Purity: Serum
Quantity: 50 µl
recommended dilution: 1 : 120 (IG), 1 : 2500-1 : 5000 (WB)
Reconstitution: For reconstitution add 50 µl of sterile water.
storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
tested applications: Immunogold (IG), Western blot (WB)
additional information: This antibody can be used as a marker for the soluble and insoluble fractions Rühle et al. (2021).

Can be provided with 0.1% ProClin, please inquire.
additional information (application): This antibody can be used to identify di and monomeric cytb6-f complexes  Kaňa et al. 2020.

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.

All references: Rühle et al. (2021) PGRL2 triggers degradation of PGR5 in the absence of PGRL1. Nat Commun. 2021 Jun 24;12(1):3941. doi: 10.1038/s41467-021-24107-7. PMID: 34168134; PMCID: PMC8225790.
von Bismarck et al. (2021) Light acclimation interacts with thylakoid ion transport to govern the dynamics of photosynthesis. Research Square; 2021. DOI: 10.21203/
Wang et al. (2020) Rerouting of ribosomal proteins into splicing in plant organelles. BioRxiv, DOI: 10.1101/2020.03.03.974766 .BN-PAGE
Simakawa et al. (2020). Near-infrared in Vivo Measurements of Photosystem I and Its Lumenal Electron Donors With a Recently Developed Spectrophotometer. Photosynth Res. , 144 (1), 63-72
Ka?a et al. (2020). Fast Diffusion of the Unassembled PetC1-GFP Protein in the Cyanobacterial Thylakoid Membrane. Life (Basel). 2020 Dec 29;11(1):E15. doi: 10.3390/life11010015. PMID: 33383642.
Aznar et al. (2020). Psb35 protein stabilizes the CP47 assembly module and associated high-light inducible proteins during the biogenesis of photosystem II in the cyanobacterium synechocystis sp. PCC6803. Plant Cell Physiol. 2020 Dec 1:pcaa148. doi: 10.1093/pcp/pcaa148. Epub ahead of print. PMID: 33258963.
Patir-Nebioglu et al. (2019). Pyrophosphate modulates plant stress responses via SUMOylation. Elife. 2019 Feb 20;8. pii: e44213. doi: 10.7554/eLife.44213.
Patil et al. (2018). FZL is primarily localized to the inner chloroplast membrane however influences thylakoid maintenance. Plant Mol Biol. 2018 Jul;97(4-5):421-433. doi: 10.1007/s11103-018-0748-3.
Giovanardi et al. (2018). In pea stipules a functional photosynthetic electron flow occurs despite a reduced dynamicity of LHCII association with photosystems. Biochim Biophys Acta. 2018 May 24. pii: S0005-2728(18)30129-4. doi: 10.1016/j.bbabio.2018.05.013.
Rantala and Tikkanen et al. (2018). Phosphorylation?induced lateral rearrangements of thylakoid protein complexes upon light acclimation. Plant Direct Vol. 2, Issue 2.
Heinnickel et al. (2016). Tetratricopeptide repeat protein protects photosystem I from oxidative disruption during assembly. Proc Natl Acad Sci U S A. 2016 Mar 8;113(10):2774-9. doi: 10.1073/pnas.1524040113
Pavlovi? et al. (2016). Light-induced gradual activation of photosystem II in dark-grown Norway spruce seedlings. Biochim Biophys Acta. 2016 Feb 18. pii: S0005-2728(16)30028-7. doi: 10.1016/j.bbabio.2016.02.009.
Liu and Last (2015). A land plant-specific thylakoid membrane protein contributes to photosystem II maintenance in Arabidopsis thaliana. Plant J. 2015 Jun;82(5):731-43. doi: 10.1111/tpj.12845. Epub 2015 Apr 29.
Dahal et al. (2015). Improved photosynthetic performance during severe drought in Nicotiana tabacum overexpressing a nonenergy conserving respiratory electron sink. New Phytol. 2015 May 29. doi: 10.1111/nph.13479.
Renato et al. (2014). Tomato fruit chromoplasts behave as respiratory bioenergetic organelles during ripening. Plant Physiol. 2014 Aug 14. pii: pp.114.243931. (western blot, immunogold)
Armbruster et al. (2014). Ion antiport accelerates photosynthetic acclimation in fluctuating light environments. Nat Commun. 2014 Nov 13;5:5439. doi: 10.1038/ncomms6439
Confirmed reactivity: Arabidopsis thaliana, Echinochloa crus-galli, Hordeum vulgare, Nicotiana tabacum, Panicum miliaceum, Picea abies, Pisum sativum, Thermosynechococcus elongatus, Solanum lycopersicum, Synechocystis sp. PCC6803, Zea mays
predicted reactivity: Brachypodium distachyon, Cannabis sativa, Cucumis melo, Oryza sativa, Populus trichocarpa, Solanum tuberosum, Sorghum bicolor, Triticum aestivum
Species of your interest not listed? Contact us
not reactive in: No confirmed exceptions from predicted reactivity are currently known.

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