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UGPase | UDP-glucose pyrophosphorylase (cytoplasm marker)

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AS05 086  |  clonality: polyclonal  |  host: rabbit  |  reactivity: A. thaliana, C. annuum, C. sativus, F. arundinacea,  H. vulgare, L. esculentum, L. chilense, Malus x domestica Borkh. c.v. Fuji, M. polymorpha, Medicago truncatula, N. tabacum, O. sativa, P. glauca, Populus sp., S. tuberosum, S. sogarandinum, Triticum |  cellular [compartment marker] of cytoplasm

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AS05 086

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product information
Background

UDP-glucose pyrophosphorylase (UGPase, UDPGP) E.C=2.7.7.9.  is a key enzyme of synthesis of sucrose, cellulose and other saccharides. There are two cytoplasmic isoforms of UGPase-A (which share 94 % identity on amino acid level) and one chloroplastic UGPase-B isoform in Arabidopsis thaliana which share ca. 10-11 % of identity (Kleczkowski et al. 2011).

Immunogen

Recombinant UGPase Q43772 overexpressed and purified from E.coli

Host Rabbit
Clonality Polyclonal
Clone
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 Immunolocalization (IL), Western blot (WB)
Related products

AS14 2813 | Anti-UDP-glucose pyrophosphorylase (cytoplasm marker) (Hordeum vulgare
For cytoplasmic marker for Chlamydomonas reinhardtii - please check NAB1

Collection of antibodies to carbohydrate metabolism

Recommended secondary antibody

Plant protein extraction buffer

Secondary antibodies

Additional information

Cellular [compartment marker] of cytoplasm, UGPse is a cytoplasmic protein Martz et al. (2002)

application information
Recommended dilution 1 : 1500 (IL), 1 : 1000-1 : 3000 (WB)
Expected | apparent MW

51.6 kDa

Confirmed reactivity Arabidopsis thaliana, Arabidopsis halleri, Brassica rapa, Capsicum annuum, Cucumis sativus, Festuca arundinacea, Hordeum vulgare, Lycopersicum esculentum, Lycopersicum chilense, Malus x domestica Borkh. c.v. Fuji,, Marchantia polymorpha, Medicago truncatula, Mesemryanthenum crystallinum, M.vaginalis, Nicotiana benthamiana,  Nicotiana tabacum, Oryza sativa, Phaseolus vulgaris, Picea glauca, Populus sp., Solanum lycopersicum, Solanum tuberosum, Solanum sogarandinu, Triticum aestivum
Predicted reactivity Amorpha fruticosa, Bambusa oldhamii, Brachypodium distachyon, Brassica pekinensis, Capsella rubella, Citrus sinensis, Cucumis melo, Eucalyptus grandis, Glycine max, Glycine soja, Gossipium hirsutum, Jatropha curcas, Pinus taeda, Populus tremula, Ricinus communis, Saccharum officinarum, Sorghum bicolor, Theobroma cacao, Zea mays, Vitis vinifera
Not reactive in

C. merolae, diatoms

Additional information

This antibody detectes 1 ng of UGPase in a western blot and reacts with both cytosolic isoforms only which have similar MW of ca. 52 kDa in Arabidopsis thaliana

Selected references Vincent et al. (2017). A genome-scale analysis of mRNAs targeting to plant mitochondria: upstream AUGs in 5' untranslated regions reduce mitochondrial association. Plant J. 2017 Dec;92(6):1132-1142. doi: 10.1111/tpj.13749.
Nagel et al. (2017). Arabidopsis SH3P2 is an ubiquitin-binding protein that functions together with ESCRT-I and the deubiquitylating enzyme AMSH3. Proc Natl Acad Sci U S A. 2017 Aug 7. pii: 201710866. doi: 10.1073/pnas.1710866114.
Schalk et al. (2017). Small RNA-mediated repair of UV-induced DNA lesions by the DNA DAMAGE-BINDING PROTEIN 2 and ARGONAUTE 1. Proc Natl Acad Sci U S A. 2017 Mar 21. pii: 201618834. doi: 10.1073/pnas.1618834114.
Castellano et al. (2016). A pathogenic long noncoding RNA redesigns the epigenetic landscape of the infected cells by subverting host Histone Deacetylase 6 activity. New Phytol. 2016 Sep;211(4):1311-22. doi: 10.1111/nph.14001. Epub 2016 May 12.
Hsu et al. (2016). Super-resolution ribosome profiling reveals unannotated translation events in Arabidopsis. Proc Natl Acad Sci U S A. 2016 Oct 21. pii: 201614788.
Liu et al. (2016). iTRAQ-based quantitative proteomic analysis reveals the role of the tonoplast in fruit senescence. J Proteomics. 2016 Sep 2;146:80-9. doi: 10.1016/j.jprot.2016.06.031.

UGPase

 A 1-year-old greehouse grown plant was dissectedinto different tissues, which were then used for enzyme assays and immunoblot analyses. Equal amounts of total protein (7.5 μg) were loaded on each lane. SDS-PAGE was run on a 7.5% gel. Immunoblot was done using Amersham PVDF transfer membrane. Primary antibodies against barley UGPase were used in 1: 1000 dilution. Secondary antibodies (Rabbit IgG, HRP-Linked Whole Antibody from donkey) were used at 1:10 000.

ff - female flower, mf - male flower, yl - young leaf, ml - mature leaf, sbk - stem bark, sph - stem phloem and cambium, sxy - stem xylem, rxy - root xylem

 

western blot using plant anti-UGPase antibodies

15 µg of  total soluble protein  extract from leaves and stems of  Solanum tuberosum (1),  Solanum sogarandinum (2),   Lycopersicum esculentum (3)Lycopersicum chilense  (4) , Arabidopsis thaliana (5) , Cucumis sativus (6)Festuca arundinacea  (7) , Nicotiana tabacum (8) and Capsicum annuum (9)  were separated  on 10% SDS-PAGE and blotted onto nitrocellulose .  After blocking with 5% milk in TBST , blots were incubated with the primary antibody at a dilution of 1:1500 in TBST for 1h  at room temperature. Following incubation and wash steps, blots were incubated with secondary Anti-Rabbit IgG , Alkaline Phosphatase Conjugate for 1 hour at a dilution of 1:40000 . Blots were developed with the alkaline phosphatase detection system using NBT/BCIP (SIGMA).

Courtesy of Bartosz Szabala, Institute of Plant Genetics, Polish Academy of Science .


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