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
Immunogen
KLH-conjugated peptide derived from known PRK sequences including Arabidopsis thaliana UniProt: P25697, TAIR: At1g32060
Host
Rabbit
Clonality
Polyclonal
Purity
Affinity purified serum in PBS, pH 7.4
Format
Lyophilized
Quantity
50 µg
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
Western blot (WB)
Recommended dilution
1 : 1000 (WB)
Expected | apparent MW
44 | 39 kDa (A. thaliana)
Reactivity
Confirmed reactivity
Arabidopsis thaliana, Chlamydomonas reinhardtii, Hordeum vulgare, Synechocystis sp. PCC6803, Synechococcus PCC 7942, Thalassiosira pseudonana, Oryza sativa, Zea mays
Predicted reactivity
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
Not reactive in
Proteobacteria
Application examples
Application examples
Applicaton example
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
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
Additional information
Additional information
Antibody can be used as a marker of chloroplast stroma.
Antibody detects PRK using a load from 4-20 µg/well of a chloroplast fraction, incubation over night at 4°C.
Background
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
Selected references
Fukayama et al. (2018). Expression level of Rubisco activase negatively correlates with Rubisco content in transgenic rice. Photosynth Res. 2018 May 30. doi: 10.1007/s11120-018-0525-9.
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.
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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