GLN2 | GS2, chloroplastic form of glutamine synthetase
AS08 296 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, N. tabacum, O. sativa, P. sativum, S. oleracea
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KLH-conjugated synthetic peptide which is a part of part of the glutamine synthetase/guanido kinase superfamily catalytic region chosen from various available sequences, including Arabidopsis thaliana GLN2, UniProt: Q43127, TAIR: AT5G35630
47 | 44-45 kDa
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0.5 µg of protein from Arabidopsis thaliana total leaf fraction (1), 5 µg of protein fromSpinacia oleracea chlorplast enriched fraction (2), molecular weight markers (MagicMarkTM,Invitrogen) (M), the same samples as in 1 and 2 but after peptide neutralisation assay, e.g. incubation of the antibody with 100 mM excess of peptide used to elicit andt-GLN2 antibody (4,5), extracted with PEB (AS08 300), were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF (Millipore). Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-GLN2 antibody (AS08 296, 1:5 000, 1h) and secondary anti-rabbit (1:20000, 1 h) antibody (HRP conjugated) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T. All steps were performed at RT with agitation. Blots were developed for 5 min with chemiluminescent detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).
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Glutamine synthetase (GLN or GS) is one of the key enzymes involved in nitrogen metabolism of plants. It catalyses the synthesis of glutamine from glutamate and ammonia in an ATP-dependent reaction. There are two general classes of glutamine synthetase in plants: GLN1, a cytosolic form and GLN2, a chloroplastic form. GLN2 is encoded by a single gene and is highly abundant in mesophyll cells of leaves for the assimilation of ammonia produced from photorespiration and the reduction of nitrate in the chloroplasts. GLN2 is a target for thioredoxin.
Chen et al. (2018). TIC236 links the outer and inner membrane translocons of the chloroplast. Nature. 2018 Dec;564(7734):125-129. doi: 10.1038/s41586-018-0713-y.
Tamburino et al. (2017). Chloroplast proteome response to drought stress and recovery in tomato (Solanum lycopersicum L.). BMC Plant Biol. 2017 Feb 10;17(1):40. doi: 10.1186/s12870-017-0971-0.
Dixit (2015). Sulfur alleviates arsenic toxicity by reducing its accumulation and modulating proteome, amino acids and thiol metabolism in rice leaves. Sci Rep. 2015 Nov 10;5:16205. doi: 10.1038/srep16205.
Lee et al. (2013). Stromal protein degradation is incomplete in Arabidopsis thaliana autophagy mutants undergoing natural senescence. BMC Res Notes, Jan 17.
Hu and Li (2012). The amino-terminal domain of chloroplast Hsp93 is important for its membrane association and functions in vivo. Plant Physiol. Apr;158(4):1656-65. doi: 10.1104/pp.112.193300. Epub 2012 Feb 21.
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