PC | Plastocyanin

AS06 141 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, B. juncea, H. annuus, L. sativus, N. tabacum, O. sativa, P. sativum, S. oleracea, S. tuberosum, Synechocystis sp. PCC6803, Z. mays

compartment marker of chloroplast thylakoid lumen

Product Information

Immunogen

Purified native plastocyanin from Spinacia oleracea UniProt: P00289

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 50 µl

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 the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.

Tested applications Immunogold (IG), Western blot (WB)

Recommended dilution 1 : 100 (IG), 1 : 2000 (WB)

Expected | apparent MW

10 kDa

Reactivity

Confirmed reactivity Arabidops thaliana, Brassica juncea, Heliantus annuus, Hordeum vulgare, Lathyrus sativus, Nicotiana tabacum, Oryza sativa, Pisum sativum, Spinacia oleracea, Solanum tuberosum, Synechocystis sp. PCC6803, Zea mays

Predicted reactivity

Catalpa bungei, Dicots, Chlamydomonas reinhardtii, Nicotiana benthamiana, Physcomitrium patens, Ricinus communis, Solanum lycopersicum

Species of your interest not listed? Contact us

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

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

western blot image using anti-plastocyanin antibody
10 µg of total protein from Arabidopsis thaliana (1), Brassica juncea (2), Zea mays (3), Oryza sativa (4), Solamum lycopersicum (5), Nicotiana tabacum (6), Heliantus annuus (7) were separated on SDS-PAGE and blotted to nitrocellulose. Filters were probed with anti-PC antibody (AS06 141, 1:2000). Signal was developed using alkaline phosphatase conjugated secondary antibody. Each sample was run in duplicate. Signal was developed using alkaline conjugated secondary antibody.

This antibody will also work well with HRP-conjugated secondary antibodies, as AS09 602.

Reactant: Malus domestica (Apple)

Application: Western Blotting

Pudmed ID: 23886449

Journal: Plant Methods

Figure Number: 2C

Published Date: 2013-07-28

First Author: Sikorskaite, S., Rajamäki, M. L., et al.

Impact Factor: 5.139

Open Publication

Analysis of protein fractions from different steps of nuclei isolation. The protein fractions obtained following the main steps of the procedure used for isolation of nuclei from leaves of apple are shown. Equal amounts of proteins (4 ?g) from each fraction were loaded on the gel. A) Lumenal-binding protein 2 (BiP2), B) histone H3 and C) plastocyanin (PC) were detected with specific antibodies by western blot analysis. D) Coomassie blue staining of the fractionated proteins separated by polyacrylamide gel electrophoresis. Lane 1: crude extract of proteins from homogenized leaf tissue; lane 2: resuspended pellet of the whole cell lysate obtained following treatment with Triton X-100 and centrifugation; lane 3: nuclei collected from the 60% Percoll layer (see nuclear fraction in Figure 1B); lane 4: the interface fraction of 60% Percoll and 2.5 M sucrose layers in the density gradient containing chloroplasts and unbroken cells (see 60% P/2.5 M S interface in Figure 1B); lane 5: sucrose layer of the density gradient (see 2.5 M sucrose layer in Figure 1B); lane 6: nuclei purified by centrifugation on a 35% Percoll cushion.

Reactant: Pisum sativum (Pea)

Application: Western Blotting

Pudmed ID: 33445673

Journal: Int J Mol Sci

Figure Number: 6A

Published Date: 2021-01-12

First Author: Tokarz, K. M., Weso?owski, W., et al.

Impact Factor: 5.542

Open Publication

Content of (a) plastocyanin (PC) and (b) large subunit of RuBisCo (RbcL) of grass pea leaf and stem photosynthetic apparatus under salinity; content of proteins expressed as relative units [RU]; different letters—statistically significant differences within each organ (leaf-uppercase, stem-lowercase) at p ? 0.05; (n = 3).

Additional information

Cellular [compartment marker] of chloroplast thylakoid lumen

This product can be sold containing ProClin if requested.

Plastocyanin runs abberant due to negative charge at 12-19 kDa on SDS-PAGE depending upon the system used. in 15 % gel the protein will run closer to its true MW than in 12 % gel. In some cases PC can be very acidic and run at twice of its MW.

PC1 runs closer to 14 kDa while PC2 runs closer to 19 kDa. For good resolution adding fresh DTT to the sample buffer is recommended.
PC2 is generally more abundant and it increases with Cu feeding. PC1 is expressed first after etiolated seedlings are placed in the light.

Related products

AS06 202 | Anti-cytc6 marker antibody of thylakoid lumen for Chlamydomonas reinhardtii, rabbit antibodies

Plant protein extraction buffer

Secondary antibodies

Background

Plastocyanin (PC) is a small Cu protein and a mobile electron carrier in the lumen of the thylkoids. PC interacts with the B/F complex and Photosystem I. Alternative name: DNA-damage-repair/toleration protein DRT112.

Product citations

Selected references Tokarz et al. (2021). Stem Photosynthesis-A Key Element of Grass Pea (Lathyrus sativus L.) Acclimatisation to Salinity. Int J Mol Sci. 2021 Jan 12;22(2):685. doi: 10.3390/ijms22020685. PMID: 33445673; PMCID: PMC7828162.
Viola et al. (2021) In vivo electron donation from plastocyanin and cytochrome c6 to PSI in Synechocystis sp. PCC6803. Biochim Biophys Acta Bioenerg. 2021 May 15;1862(9):148449. doi: 10.1016/j.bbabio.2021.148449. Epub ahead of print. PMID: 34004195.
Furutani et al. (2021) The difficulty of estimating the electron transport rate at photosystem I. J Plant Res. 2021 Nov 15. doi: 10.1007/s10265-021-01357-6. Epub ahead of print. PMID: 34778922.
Wang et al. (2020) Rerouting of ribosomal proteins into splicing in plant organelles. BioRxiv, DOI: 10.1101/2020.03.03.974766 .
Galvis et al. (2020). H+ transport by K+ EXCHANGE ANTIPORTER3 promotes photosynthesis and growth in chloroplast ATP synthase mutants. Plant Physiol. pp.01561.2019. doi: 10.1104/pp.19.01561.
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
Cha et al. (2019). Arabidopsis GIGANTEA negatively regulates chloroplast biogenesis and resistance to herbicide butafenacil. Plant Cell Rep. 2019 Jul;38(7):793-801. doi: 10.1007/s00299-019-02409-x.
Mermod et al. (2019). SQUAMOSA promoter-binding protein-like 7 mediates copper deficiency response in the presence of high nitrogen in Arabidopsis thaliana. Plant Cell Rep. 2019 May 15. doi: 10.1007/s00299-019-02422-0.
Balyan et al. (2017). Identification of miRNA-mediated drought responsive multi-tiered regulatory network in drought tolerant rice, Nagina 22. Sci Rep. 2017 Nov 13;7(1):15446. doi: 10.1038/s41598-017-15450-1.
Perea-García et al. (2017). Arabidopsis copper transport protein COPT2 participates in the cross talk between iron deficiency responses and low-phosphate signaling. Plant Physiol. 2013 May;162(1):180-94. doi: 10.1104/pp.112.212407.
Yoshida et al. (2016). Hisabori T1.Two distinct redox cascades cooperatively regulate chloroplast functions and sustain plant viability. Proc Natl Acad Sci U S A. 2016 Jul 5;113(27):E3967-76. doi: 10.1073/pnas.1604101113. Epub 2016 Jun 22.
Kropat et al. (2015). Copper economy in Chlamydomonas: Prioritized allocation and reallocation of copper to respiration vs. photosynthesis. Proc Natl Acad Sci U S A. 2015 Feb 2. pii: 201422492.
Sook Seok et al. (2013). AtFKBP16-1, a chloroplast lumenal immunophilin, mediates response to photosynthetic stress by regulating PsaL stability. Physiologia Plantarum, DOI: 10.1111/ppl.12116.
Perera-Garcia et al. (2013). Arabidopsis copper transport protein COPT2 participates in the crosstalk between iron deficiency responses and low phosphate signaling.