APX | L-ascorbate peroxidase
AS08 368 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, A.maritima, C. annuum, Citrus sp., D. sanguinalis, E. crus-galli, l. formosana, M. esculenta, M. sativa, N.tabacum, O. sativa, P. miliaceum, S. oleracea, S. superba, S. vulgaris, Triticum aestivum, Z. mays
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Product Information
Synthetic peptide derived from Arabidopsis thaliana tAPX (acession At1g77490) and sAPX (At4g08390) protein sequences, coupled to BSA. Five out of twelve amino acids are also identical with cAPX1 (At1g07890), cPX2 (At3g09640) and pAPX (At4g35000)
25-38 kDa for A. thaliana
Reactivity
Brassica rapa subsp. oleifera Stromal APX; Glycine max, Glycine soja L-ascorbate peroxidase T, chloroplastic; Medicago truncatula thylakoid-bound APX; Pisum sativum Chloroplast stromal ascorbate peroxidase 12; Solanum lycopersicum thylakoid-bound APX; Spinacia oleracea stromal APX; Theobroma cacao L-APX T isoform 3; Vitis vinifera
Application examples
Application example
5 to 20 μg of total leaf protein from Arabidopsis thaliana (left panel) and chloroplast fractions (thylakoids and soluble, right panel) was separated on 15% polyacrylamide gel with 6M urea and blotted on PVDF. Filters were blocked 1h with 5% BSA, incubated with anti-APX antibody (1: 2000, 1h) followed by incubation with secondary HRP-coupled anti rabbit antibody (1: 10 000, 1h). Signal was detected with chemiluminescence detection reagent. AS08 368 is reactive to thylakoid (tAPX, 38 kDa), stromal (sAPX, 33 kDa), peroxisomal (pAPX, 31 kDa) and cytoplasmic (cAPX1 + cAPX2, 25 kDa) forms of ascorbate peroxidases.
Total proteins of Arabidopsis thaliana leaves were extracted with 10 % TCA and precipitated. The pellet was washed with acetone and resuspended in 100mM Tris-HCl (pH 7.5), 1mM EDTA, 2% (w= v) SDS, 1:100 of protease inhibitor cocktail (Thermo Scientific), 1 mM PMSF. Leaves were also grinded in 100 mM Tris-HCl (pH 7.5), MgCl2 10 mM, 1 mM EDTA, 1 mM PMSF, 1/100 of protease inhibitor cocktail and centrifugated. The supernatant (soluble fraction) was separated and the pellet (membrane fraction) was resuspended in the same buffer with 6 M urea and 1% SDS. Different amounts of proteins were separated in 15 % polyacrylamide gel with 6M urea after denaturation (70°C 5 min) and blotted on PVDF. Filters were blocked 1h with 5% BSA, Incubated with anti-APX antibodies at a dilution 1:2000, 1h/RT, washed 4 times with TBS tween (5 min each) and incubated with HRP coupled anti-rabbit IgG secondary antibody in dilution 1:10 000 1h/RT (AS09 602, Agrisera). After incubation with secondary antibody, the filter was washed 4 times with TBS (5 min each) and signal was detected with chemiluminescent detection reagent (30 secs exposition in film).
Courtesy Manuel Guinea Diaz, University of Turku, Finland
Additional information
This product can be sold containing proclin if requested
Background
APX plays a key role in plant antioxidant system by reducing hydrogen peroxide to water. Cellular localization includes chloroplast (tAPX and sAPX), cytosol (cAPX) and peroxisome (pAPX).
Product citations
Deng et al. (2019). Integrated proteome analyses of wheat glume and awn reveal central drought response proteins under water deficit conditions. J Plant Physiol. 2019 Jan;232:270-283. doi: 10.1016/j.jplph.2018.11.011.
Balfagón et al. (2018). Involvement of ascorbate peroxidase and heat shock proteins on citrus tolerance to combined conditions of drought and high temperatures. Plant Physiol Biochem. 2018
Cunha et al. (2016). Salinity and osmotic stress trigger different antioxidant responses related to cytosolic ascorbate peroxidase knockdown in rice roots. Environmental and Experimental Botany, Volume 131, November 2016, Pages 58-67.
Ko et al. (2016). Constitutive expression of a fungus-inducible carboxylesterase improves disease resistance in transgenic pepper plants. Planta. 2016 Aug; 244(2):379-92. doi: 10.1007/s00425-016-2514-6. Epub 2016 Apr 13.
Yin et al. (2016). Comprehensive Mitochondrial Metabolic Shift during the Critical Node of Seed Ageing in Rice. PLoS One. 2016 Apr 28;11(4):e0148013. doi: 10.1371/journal.pone.0148013. eCollection 2016.
Vuleta et al. (2016). Adaptive flexibility of enzymatic antioxidants SOD, APX and CAT to high light stress: The clonal perennial monocot Iris pumila as a study case. Plant Physiol Biochem. 2016 Mar;100:166-73. doi: 10.1016/j.plaphy.2016.01.011. Epub 2016 Jan 19
Hattab et al. (2015). Characterisation of lead-induced stress molecular biomarkers in Medicago sativa plants. Environm. Exp. Botany. Volume 123, March 2016, Pages 1–12.
Parys et al. (2014). Metabolic Responses to Lead of Metallicolous and Nonmetallicolous Populations of Armeria maritima. Arch Environ Contam Toxicol. 2014 Jul 29.
Feifei et al. (2014). Comparison of Leaf Proteomes of Cassava (Manihot esculenta Crantz) Cultivar NZ199 Diploid and Autotetraploid Genotypes. PLoS One. 2014 Apr 11;9(4):e85991. doi: 10.1371/journal.pone.0085991. eCollection 2014.
Sobrino-Plata et al. (2014). Glutathione is a key antioxidant metabolite to cope with mercury and cadmium stress. Plant Soil, DOI 10.1007/s11104-013-2006-4.
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