PRK ribulose-5-P-kinase | Phosphoribulokinase

PRODUCT INFORMATION IN PDF

product 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 Immunogen KLH-conjugated peptide derived from known PRK sequences including Arabidopsis thaliana P25697, At1g32060 Host Rabbit Clonality Polyclonal Clone Purity Affinity purified serum in PBS, pH 7.4 Format Lyophilized in PBS pH 7.4 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) Related products collection of antibodies to various photosynthetic proteins Additional information Antibody can be used as a marker of chloroplast stroma.

application information
Recommended dilution		1 : 1000 (WB)
Expected | apparent MW		44 | 39 kDa (A. thaliana)
Confirmed reactivity		Arabidopsis thaliana, Chlamydomonas reinhardtii, Hordeum vulgare, Synechocystis sp. PCC6803, Synechococcus PCC 7942, Thalassiosira pseudonana, Zea mays
Predicted reactivity		Glycine max, Hordeum vulgare, Malus domestica, Micromonas sp., Ostreocossus tauri, Oryza sativa, Physcomitrella patens, Populus trichocarpa, Spinacia oleracea, Solanum tuberosum,  Sorghum bicolor,  Synechocystis PCC 6803, Synechococcus elongatus, Zea mays, Vitis vinifera
Not reactive in		Proteobacteria
Additional information		Antibody detects PRK using a load from 4-20 µg/well of a chloroplast fraction, incubation ON at 4°C.
Selected references		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)

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 ECL according to the manufacturer's instructions.

Courtesy of Lauri Nikkanen, University of Turku, Finland

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