PRK ribulose-5-P-kinase | Phosphoribulokinase
AS07 257 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, H. vulgare, O. sativa, Z. mays, cyanobacteria, diatoms
compartment marker of chloroplast stroma
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Product Information
KLH-conjugated peptide derived from known PRK sequences including Arabidopsis thaliana UniProt: P25697, TAIR: At1g32060
44 | 39 kDa (A. thaliana)
Reactivity
Glycine max, Hordeum vulgare, Lactuca sativa, Malus domestica, Micromonas sp., Ostreocossus tauri, Physcomitrium patens, Populus trichocarpa, Spinacia oleracea, Solanum tuberosum, Sorghum bicolor, Synechocystis PCC 6803, Synechococcus elongatus, Zea mays, Vitis vinifera
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Application examples
WT-Col-0 Arabidopsis thaliana leaves were frozen in liquid nitrogen and soluble proteins were extracted in a buffer containing 50 mM HEPES, 5 mM NaCl and 10 mM MgCl2. 10 µg, 20 or 40 ug of eparated in SDS-PAGE (15% acrylamide with 6M urea) and blotted 1h to a PVDF membrane using Höefer semi-dry blotter. Blot was blocked with 4 % milk in TTBS for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 overnight in +4°C. The antibody solution was decanted and the blot was washed 3 x 5 min with TTBS at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:20 000 in 1% milk/TTBS for 2h at RT with agitation. The blot was washed 3x5 min in TTBS and 1x5 min in TBS at RT with agitation and developed for 5 min with chemiluminescent detection reagent, according to the manufacturer's instructions.
Courtesy of Lauri Nikkanen, University of Turku, Finland
Reactant: Arabidopsis thaliana (Thale cress)
Application: Western Blotting
Pudmed ID: 32772439
Journal: Plant J
Figure Number: 4A
Published Date: 2020-11-01
First Author: Guinea Diaz, M., Nikkanen, L., et al.
Impact Factor: 5.526
Open PublicationRedox states of the ??subunit of ATP synthase (CF1?), phosphoribulokinase (PRK) and 2?Cys peroxiredoxins (2?Cys PRX) in dark?adapted plants and plants illuminated under various irradiances.Leaves from 35?day?old Col?0, ntrc and transgenic lines overexpressing (OE) modified TRX systems (OE?NTRC, OE?NTR?TRXf, and OE?SGPS) were collected at the end of the night (D), or from plants illuminated for 2 h at 50 (low light, LL), 200 (growth light, GL) and 800 (high light, HL) ?mol photons m?2 sec?1 and frozen in liquid nitrogen. Total leaf proteins were extracted and separated in sodium dodecyl sulphate–polyacrylamide gel electrophoresis as described in Figure 1 before Western blotting with anti?2?Cys Prx antibody, anti?Cf1? antibody and anti?PRK antibody. Bands corresponding to the oxidized (Ox) and reduced (Red) form of the proteins are marked in the figure. 2?Cys PRX exist as three redox forms, fully oxidized (Ox), partially reduced (1SH) and fully reduced (2SH) (Nikkanen et al., 2016). Ponceau staining was used as the loading control.
Additional information
Background
PRK ribulose-5-P-kinase| phosphoribulokinase is an enzyme that catalyzes the chemical reaction ATP + D-ribulose 5-phosphate to ADP + D-ribulose 1,5-bisphosphate.
Product citations
Pérez-Ruiz et al. (2017). NTRC-dependent redox balance of 2-Cys peroxiredoxins is needed for optimal function of the photosynthetic apparatus. Proc Natl Acad Sci U S A. 2017 Nov 7;114(45):12069-12074. doi: 10.1073/pnas.1706003114.
Rai et al. (2017). Real-time iTRAQ-based proteome profiling revealed the central metabolism involved in nitrogen starvation induced lipid accumulation in microalgae. Sci Rep. 2017 Apr 5;7:45732. doi: 10.1038/srep45732. (microalga, western blot)
Nikkanen et al. (2016). Crosstalk between chloroplast thioredoxin systems in regulation of photosynthesis. Plant Cell Environ. 2016 Aug;39(8):1691-705. doi: 10.1111/pce.12718.
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