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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44 | 39 kDa (A. thaliana)
Glycine max, Hordeum vulgare, Malus domestica, Micromonas sp., Ostreocossus tauri, Physcomitrella patens, Populus trichocarpa, Spinacia oleracea, Solanum tuberosum, Sorghum bicolor, Synechocystis PCC 6803, Synechococcus elongatus, Zea mays, Vitis vinifera
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 ECL according to the manufacturer's instructions.
Courtesy of Lauri Nikkanen, University of Turku, Finland
Antibody can be used as a marker of chloroplast stroma.
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
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)