UAGPase | UDP-GlcNAc pyrophosphorylase
AS14 2829 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, H. vulgare, Nicotiana tabacum
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Product Information
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 µl
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 : 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
Species of your interest not listed? Contact us
Not reactive in
No confirmed exceptions from predicted reactivity are currently known.
Application examples
Application examples
Application example
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.
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
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.
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.
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