PC | Plastocyanin
AS06 141 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, B. juncea, H. annuus, N. tabacum, O. sativa, P. sativum, S. oleracea, S. tuberosum, Z. mays | cellular [compartment marker] of chloroplast thylakoid lumen
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Purified native plastocyanin from Spinacia oleracea UniProt: P00289
Catalpa bungei, Dicots, Physcomitrella patens, Ricinus communis, Solanum lycopersicum
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
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.
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.
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.
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.