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UGPase | UDP-glucose pyrophosphorylase (cytoplasm marker) (Hordeum vulgare)

AS14 2813  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A. thaliana, H.vulagre, Z.mays

UGPase | UDP-glucose pyrophosphorylase (cytoplasm marker) (Hordeum vulgare) in the group Antibodies Plant/Algal  / Carbohydrates at Agrisera AB (Antibodies for research) (AS14 2813)
UGPase | UDP-glucose pyrophosphorylase (cytoplasm marker) (Hordeum vulgare)



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Product Information

Immunogen

His-tagged, full length Hordeum vulgare UGPase, overexpressed and purified from E.coli, UniProt: Q43772.1

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl of sterile water
Storage Store lyophilized/reconstituted at -20°C; 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

52 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Hordeum vulgare, Zea mays
Predicted reactivity

Bambusa oldhamii, Brassica pekinensis, Brassica rapa, Capsicum annuum, Cucumis sativus, Dendrobium catenatum, Dendrocalamus sinicus,  Glycine max, Gossipium hirsutum, Lycopersicum esculentum, Lycopersicum chilense, Marchantia polymorpha, Oryza sativa, Picea glauca, Populus sp., Solanum tuberosum, Populus tremula, Ricinus communis, Saccharum officinarum, Vitis vinifera, for more species, please Species of your interest not listed? inquire


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

western blot using anti-barley UGPase antibodies
10 µg of total protein from Arabidopsis thaliana leaf (1) , Hordeum vulgare leaf (2)Zea mays leaf (3), recombinant UGPase 0.5 pmol (4),  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 (GE RPN 2125; Healthcare) 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.

Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 27109828

Journal: Nat Commun

Figure Number: 6A

Published Date: 2016-04-25

First Author: Yang, B. J., Han, X. X., et al.

Impact Factor: 13.783

Open Publication

Auxin stimulates PTRE1 accumulation at the plasma membrane.(a) Western blot analysis showed that auxin treatment results in relatively more PTRE1 at the plasma membrane, but decreased levels in nucleus and cytoplasm. Arabidopsis seedlings expressing PTRE1-GFP were treated with NAA (1??M, 60 or 120?min) and then surface-exposed membrane proteins were analysed using PTRE1 antibody or plasma membrane marker H+-ATPase. Nucleus and cytoplasm fractions were prepared from Col seedlings and analysed by western blot using antibody against PTRE1, nuclear marker H3 or cytoplasm marker UGPase. The relative quantities of the proteins were calculated by using image pro plus and indicated. (b) A proposed model for how PTRE1 and TIR1 coordinate auxin responses regulating proteasome activity and Aux/IAA protein degradation. Under normal condition (with basal auxin levels), 26S proteasome activity is maintained by appropriate distribution of PTRE1 at the plasma membrane and in intracellular compartments. In response to auxin, auxin rapidly stimulates the association of TIR1 and Aux/IAA proteins resulting in degradation of Aux/IAAs (1). Later, auxin suppresses PTRE1 to inhibit proteasome activity (possibly through stimulating the accumulation of PTRE1 at plasma membrane, resulting in decreased intracellular and nuclear localization) and hence suppresses Aux/IAA protein degradation (2) to coordinate regulation of auxin responses, reflecting a mechanism for fine control of Aux/IAA homeostasis and auxin signalling.

Additional information

Additional information Cellular [compartment marker] of cytoplasm, UGPse is a cytoplasmic protein Martz et al, (2002)
This antibody is also recognizing recombinant UGPase, below 0,5 pmol

Related products

Background

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).

Alternative name: UTP--glucose-1-phosphate uridylyltransferase.

Product citations

Selected references Kleczkowski LA & Decker DD (2015) Sugar activation for production of nucleotide sugars as substrates for glycosyltransferases in plants. J. Appl. Glycosci. (in press).
immunogen:

His-tagged, full length Hordeum vulgare UGPase, overexpressed and purified from E.coli, UniProt: Q43772.1

Reconstitution: For reconstitution add 50 ĩl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 50 ĩl
storage: Store lyophilized/reconstituted at -20°C; 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)
calculated | apparent molecular mass [kDa]:

52 kDa

Confirmed reactivity: Arabidopsis thaliana, Hordeum vulgare, Zea mays
predicted reactivity:

Bambusa oldhamii, Brassica pekinensis, Brassica rapa, Capsicum annuum, Cucumis sativus, Dendrobium catenatum, Dendrocalamus sinicus,  Glycine max, Gossipium hirsutum, Lycopersicum esculentum, Lycopersicum chilense, Marchantia polymorpha, Oryza sativa, Picea glauca, Populus sp., Solanum tuberosum, Populus tremula, Ricinus communis, Saccharum officinarum, Vitis vinifera, for more species, please Species of your interest not listed? inquire


Species of your interest not listed? Contact us
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer): application example

western blot using anti-barley UGPase antibodies
10 µg of total protein from Arabidopsis thaliana leaf (1) , Hordeum vulgare leaf (2)Zea mays leaf (3), recombinant UGPase 0.5 pmol (4),  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 (GE RPN 2125; Healthcare) 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.
More images:

Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 27109828

Journal: Nat Commun

Figure Number: 6A

Published Date: 2016-04-25

First Author: Yang, B. J., Han, X. X., et al.

Impact Factor: 13.783

Open Publication

Auxin stimulates PTRE1 accumulation at the plasma membrane.(a) Western blot analysis showed that auxin treatment results in relatively more PTRE1 at the plasma membrane, but decreased levels in nucleus and cytoplasm. Arabidopsis seedlings expressing PTRE1-GFP were treated with NAA (1??M, 60 or 120?min) and then surface-exposed membrane proteins were analysed using PTRE1 antibody or plasma membrane marker H+-ATPase. Nucleus and cytoplasm fractions were prepared from Col seedlings and analysed by western blot using antibody against PTRE1, nuclear marker H3 or cytoplasm marker UGPase. The relative quantities of the proteins were calculated by using image pro plus and indicated. (b) A proposed model for how PTRE1 and TIR1 coordinate auxin responses regulating proteasome activity and Aux/IAA protein degradation. Under normal condition (with basal auxin levels), 26S proteasome activity is maintained by appropriate distribution of PTRE1 at the plasma membrane and in intracellular compartments. In response to auxin, auxin rapidly stimulates the association of TIR1 and Aux/IAA proteins resulting in degradation of Aux/IAAs (1). Later, auxin suppresses PTRE1 to inhibit proteasome activity (possibly through stimulating the accumulation of PTRE1 at plasma membrane, resulting in decreased intracellular and nuclear localization) and hence suppresses Aux/IAA protein degradation (2) to coordinate regulation of auxin responses, reflecting a mechanism for fine control of Aux/IAA homeostasis and auxin signalling.

additional information: Cellular [compartment marker] of cytoplasm, UGPse is a cytoplasmic protein Martz et al, (2002)
additional information (application): This antibody is also recognizing recombinant UGPase, below 0,5 pmol
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).

Alternative name: UTP--glucose-1-phosphate uridylyltransferase.

All references: Kleczkowski LA & Decker DD (2015) Sugar activation for production of nucleotide sugars as substrates for glycosyltransferases in plants. J. Appl. Glycosci. (in press).

Related products: UGPase | UDP-glucose pyrophosphorylase (cytoplasm marker) (Hordeum vulgare)

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