PEPC | Phosphoenolpyruvate carboxylase
AS09 458 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. comosus, A. thaliana, C. ciliaris, C. gayana, C. velia, H. vulgare, J. curcas, L. fusca, Lupinus sp. , M. maximus, M. crystallinum, N. tabacum, O. sativa, P. antidotale, P. coloratum, P. strobus, Saccharum spp. hybrid clone C91-301, S. lanata, S. laricifolia, S. bicolor, Synechocystis PCC 6803, Phaeodactylum tricornutum (strain CCAP 1055/1), T. weissfloggi, Z. mays, Z. muelleri
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KLH-conjugated synthetic peptide well conserved PEPC1 and sequences from different plant species including Arabidopsis thaliana Q9MAH0, At1g53310 (PEPC 1), Q84VW9, At3g14940 (PEPC 3). The peptide chosen to elicit this antibody is also perfectly conserved in bacterial type of this enzyme NP_177043.2 (PEPC 4).
For Zea mays, the peptide is converved in PEP1 and PEP4 isoforms.
110 | 105 kDa
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5 µg of total protein from (1) Arabidopsis thaliana leaf extracted with Protein Extration Buffer, PEB (AS08 300), (2) Spinacia oleracea total cell, extracted with PEB, (3) Hordeum vulgare total cell extracted with PEB, (4) Zea mays total cell extracted with PEB, were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% blocking reagent (GE Healthcare) 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 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:50 000 in 2% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescent detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).
10 µg of total protein extracted freshly from Arabidopsis thaliana wt leaf tissue (Atn non-senescent leaves), Arabidopsis thaliana wt leaf tissue (Ats senescent leaves), Pinus strobus needle tissue (PS0, PS36 ) with 1 M Tris-HCl, pH 6.8, 10 % SDS, 15 % sucrose, 0.5 DTT and denatured at 75°C for 5 min. were separated on 10 % Bis-Tris Nupage Novex gel (120 V/45 min. using MES buffer system) and blotted 30 min. to PVDF. Blot was blocked with 5 % non-fat milk 45 min./RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 for 1h/RT with agitation in TBS with 2 % non-fat milk or ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice for 10 min. in TBS at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:75 000 in for 1h/RT with agitation. The blot was washed as above and developed using chemiluminescent detection. Exposure time was 40 seconds.
Courtesy of Dr. Christine Yao-Yun Chang and the Ensminger lab, University of Toronto, Canada
PEPC (phosphoenolpyruvate carboxylase), EC=188.8.131.52, belongs to an enzyme family of carboxy-lyases that is catalyzing adding fo carbon dioxide to phosphoenolpyruvate (PEP) to form oxaloacetate. Alternative names: PEPCase 1, PEPCase 3, PEPC 1, PEPC 3
Durall et al. (2021). Production of succinate by engineered strains of Synechocystis PCC 6803 overexpressing phosphoenolpyruvate carboxylase and a glyoxylate shunt. Microb Cell Fact. 2021 Feb 8;20(1):39. doi: 10.1186/s12934-021-01529-y. PMID: 33557832; PMCID: PMC7871529.
Kramer et al. (2020). N6â?methyladenosine and RNA secondary structure affect transcript stability and protein abundance during systemic salt stress in Arabidopsis. Plant Direct . 2020 Jul 24;4(7):e00239.doi: 10.1002/pld3.239.
Durall et al. (2020). Increased ethylene production by overexpressing phosphoenolpyruvate carboxylase in the cyanobacterium Synechocystis PCC 6803. Biotechnol Biofuels. 2020 Jan 28;13:16. doi: 10.1186/s13068-020-1653-y.
Wang et al. (2019). PUB25 and PUB26 Promote Plant Freezing Tolerance by Degrading the Cold Signaling Negative Regulator MYB15.
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Liu et al. (2017). Plasma Membrane CRPK1-Mediated Phosphorylation of 14-3-3 Proteins Induces Their Nuclear Import to Fine-Tune CBF Signaling during Cold Response. Mol Cell. 2017 Apr 6;66(1):117-128.e5. doi: 10.1016/j.molcel.2017.02.016.
Ribeiro et al. (2017). Increased sink strength offsets the inhibitory effect of sucrose on sugarcane photosynthesis. J Plant Physiol. 2017 Jan;208:61-69. doi: 10.1016/j.jplph.2016.11.005.
Shen et al. (2016). The existence of C4-bundle-sheath-like photosynthesis in the mid-vein of C3 rice. Rice (N Y). 2016 Dec;9(1):20. doi: 10.1186/s12284-016-0094-5. Epub 2016 May 10.
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