UGPase | UDP-glucose pyrophosphorylase (cytoplasm marker)

AS05 086 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, C. annuum, C. sativus, F. margarita Swingle, 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

Product Information

Immunogen

Recombinant UGPase Q43772 overexpressed and purified from E.coli

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 Immunolocalization (IL), Western blot (WB)

Recommended dilution 1 : 1500 (IL), 1 : 1000-1 : 5000 (WB)

Expected | apparent MW

51.6 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Arabidopsis halleri, Brassica rapa, Capsicum annuum, Fortunella margarita Swingle, Citrus sinensis, 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 aestivu, Zea mays

Predicted reactivity Amorpha fruticosa, Bambusa oldhamii, Brachypodium distachyon, Brassica pekinensis, Capsella rubella, 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
Species of your interest not listed? Contact us

Not reactive in

C. merolae, diatoms

Application examples

Application examples $Western blot on cytoplasmic and chloroplastic fractions with anti-UGPase antibodies$

UGPase was used as a negative control for cytoplasm contamination in chloroplast fraction.

Total protein (first 2 lanes to the left) or chloroplast protein (2 lanes to the right) of Arabidopsis thaliana were extracted with 100 mM Tris–HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA, 0.5% Triton X-100, 1 mM DTT, 1 mM PMSF, protease inhibitor cocktail, denatured with Laemmli buffer (62.5 mM Tris–HCl pH 6.8, 2% SDS, 10% glycerol, 100 mM DTT, 0.1% β-mercaptoethanol) at 95°C 5 min and separated on 8% SDS-PAGE and blotted 1h to PVDF, using wet transfer. Blot was blocked with 5% milk for 1h/RT or 4°C/ON with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 for 1h/RT with agitation in TBS-T. The antibody solution was decanted and the blot was washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in Calbiochem matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25 000 in for 1h/RT with agitation. The blot was washed as above and developed for 10 min with Clarity Western ECL Substrate (Biorad) and visualised with chemiluminescence CCD camera Fluorchem SP (Gel Biosciences). Exposure time was 1-5 minutes.

This antibody is required to be used on chloroplast fraction. There is no signal in the total cell extract.

Courtesty of Dr. Aleksandra Kwaśnik, Warsaw University, Poland

UGPase

A 1-year-old greehouse grown plant was dissected into 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 PVDF transfer membrane. Primary antibodies against barley UGPase were used in 1: 1000 dilution. Secondary antibodies (Rabbit IgG, HRP conjugated) 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.

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

Additional information

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

This product can be sold containing ProClin if requested.

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.

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

Plant protein extraction buffer

Secondary antibodies

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).