NR | Nitrate reductase, assimilatory (100 ĩl)
AS08 310-100 | clonality: polyclonal | host: rabbit | reactivity: A. thaliana, H. vulgare, Ch. reinhardtii, red alga Gracilaria gracilis, diatom Thalassiosira sp. , P. tricornutum Bohlin, P. yunnanensis Dode, S. tuberosum
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|Recommended dilution||1 : 500-1 : 1000 (WB)|
|Expected | apparent MW||
103 kDa | 117 kDa
|Confirmed reactivity||Arabidopsis thaliana, Hordeum vulgare, Chlamydomonas reinardtii, red alga Gracilaria gracilis, diatom Thalassiosira sp., Phaeodactylum tricornutum Bohlin accession Pt1 8.6 ,Populus yunanensis Dode, Solanum tuberosum, Vigna radiata
|Predicted reactivity||Arabis alpina, Brachypodium distachyon, Brassica napus, Brassica rapa subsp. pekinensis, Capsella rubella, Citrus clementina, Citrus sinensis, Chlorella vulgaris, Dunaliella salina, marine Diatoms, Coffea canephora, Eucalyptus grandis, Glycine max, Glycine soja, Gossypium arboretum, Lycopersicum esculentum, Morus alba, Nicotiana tabacum, Nicotiana attenuata, Nicotiana benthamiana, Oryza sativa, Phaseolus vulgaris, Phytophthora infestans, Physcomitrella patens, Prunus persica, Ricinus communis, Sorghum bicolor, Spinacia oleracea, Solanum lycopersicum, Theobroma cacao, Zea mays, Vitis vinifera|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
ECL based detection systems are adviced to use since to low signal intensity can be obtaied with BCIP/NBT system.
For working with diatom samples ECL Advance (GE Healthcare) or other more sensitive ECL detection reagent is recommended.
|Selected references||Cheng et al. (2015). Quantitative proteomics analysis reveals that S-nitrosoglutathione reductase (GSNOR) and nitric oxide signaling enhance poplar defense against chilling stress. Planta. 2015 Aug 2.
Johnson and Lecomte (2015). Characterization of the truncated hemoglobin THB1 from protein extracts of Chlamydomonas reinhardtii. v1; ref status: indexed,
Zhang et al. (2014). Heterologous expression of AtPAP2 in transgenic potato influences carbon metabolism and tuber development. FEBS Lett. 2014 Aug 27. pii: S0014-5793(14)00621-8. doi: 10.1016/j.febslet.2014.08.019.
Beyzaei et al. (2014). Response of Nitrate Reductase to Exogenous Application of 5-Aminolevulinic Acid in Barley Plants. J. Plant Growth Regulation, April 2014.
Frada et al. (2013). Quantum requirements for growth and fatty acid biosynthesis in the marine diatom Phaeodactylum tricornutum (Bacilloriophyceae) in nitrogen replete and limited conditions. J. Phycology. Diatom growth and lipid efficiency
20 µg of total protein from Arabidopsis thaliana leaf (1) and Hordeum vulgare leaf (2) were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70°C for 5 min and keept on ice before loading. Protein samples were separated on 4-12% Bolt Plus gels, LDS-PAGE and blotted for 70 minutes to PVDF using tank transfer. Blots were blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) or 5% non-fat milk dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 5 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS10 1489, Agrisera) diluted to 1:20 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with TMA-6 (Lumigen) detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (VersaDoc MP 4000) and Quantity One software (Bio-Rad). Exposure time was 5 minutes.
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