UAGPase | UDP-GlcNAc pyrophosphorylase

Product no: AS14 2829

AS14 2829  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A. thaliana, H. vulgare, Nicotiana tabacum

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  • Product Info
  • Immunogen:

    His-tagged full length recombinant UAGPase of Arabidopsis thaliana, overexpressed and purified from E.coli, UniProt:O64765,, TAIR:AT2G35020. Sequence used for immunization is conserved in both isoforms: UDP-N-acetylglucosamine diphosphorylase 1 (Q940S3) and (O64765)

    Host: Rabbit
    Clonality: Polyclonal
    Purity: Serum
    Format: Lyophilized
    Quantity: 50 l
    Reconstitution: For reconstitution add 50 l of sterile water
    Storage: Store lyophilized/reconstituted at -20C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Western blot (WB)
    Recommended dilution: 1 : 10 000 (WB)
    Expected | apparent MW: 55,76 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Hordeum vulgare, Nicotiana tabacum
    Predicted reactivity: Glycne soja, Medicago truncatula, Morus notabilis, Populus trichocarpa, Theobroma cacao, Zea mays, for more species, please inquire
    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Application example

    western blot using anti-UAGPase antibodies

    10 µg of total protein from Arabidopsis thaliana leaf (1) Hordeum vulgare leaf (2), Nicotiana tabacum (3), and  recombinant UGPase 0.25 pmol (R), 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 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: 10 000 (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 AS09 602, Agrisera) diluted to 1:25 000 in blocking reagent for 1h at room temperature with agitation. The blots were washed as above. The blot was developed for 5 min with 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 10 seconds for recombinant UAGpase and 1 minute for plant extracts.
  • Additional Information
  • Additional information (application): This antibody is recognizing recombinant UAGPase at 0,25 pmol
  • Background
  • Background: UAGPase (UDP-GlcNAc pyrophosphorylase) is involved in the biosynthesis of UDP-glucosamine, an essential precursor for glycoprotein and glycolipid synthesis and is also used for regulatory protein modification in signaling pathways. The enzyme is localized in a cytoplasm. Alternative names: N-acetylglucosamine-1-phosphate uridylyltransferase 2, UDP-N-acetylgalactosamine diphosphorylase 2, UTP--glucose-1-phosphate uridylyltransferase 2, AGX.
  • Product Citations
  • Selected references: Fernández-San Millán et al. (2018). Physiological Performance of Transplastomic Tobacco Plants Overexpressing Aquaporin AQP1 into Chloroplast Membranes. J Exp. Bot. ery148, https://doi.org/10.1093/jxb/ery148.
    Kleczkowski LA & Decker DD (2015) Sugar activation for production of nucleotide sugars as substrates for glycosyltransferases in plants. J. Appl. Glycosci. 2015 Volume 62 Issue 2 Pages 25-36.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials
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AS16 ECL-S | extreme low femtogram detection

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