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
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Recombinant UGPase Q43772 overexpressed and purified from E.coli
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C. merolae, diatoms
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
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
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 .
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.
UDP-glucose pyrophosphorylase (UGPase, UDPGP) E.C=188.8.131.52. 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).
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