DHAR1 | Dehydroascorbate Reductase 1
AS11 1746 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Solanum lycopersicum
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23.6 | 23.4 kDa
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1cm2 of a leaf from Arabidopsis thaliana Col-0 (1) and or t-DNA insertion lines dhar1-1 (2), dhar1-2 (3), dhar1-3 (4), dhar2-1 (5), dhar2-2 (6), dhar1-3 EOS-DHAR1 (7), was extracted using 200µl Lyse&Load-Buffer (Grefen et al. 2009). 10 µl were separated on a 15% SDS-PAGE and blotted 1h to PVDF (using Bjerrum Buffer in a semidry blot). Blots were blocked with 5% Milk in 1xTBS-Tween20 (1%) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:5000 (in 5% Milk 1xTBS-Tween20 (1%) + 0.01 % NaN3) ON at 4°C with agitation. The antibody solution was decanted and the blot was washed 3 times for 10 minutes with 1x TBS-Tween20 at RT with agitation. Blot was incubated in secondary antibody BioRad anti-rabbit IgG AP-conjugate (#170-6518) diluted to 1:2000 in 5% Milk 1xTBS-Tween20 (1%) + 0.01 % NaN3 for 1h at RT with agitation. The blot was washed as above, equilibrated in staining buffer (100mM Tris-HCl, 100mM NaCl, 5mM MgCl2, see Grefen et al. 2009) and developed for 5-15 min. with staining solution (Nitro blue tetrazolium chloride (NBT) and 5-bromo-4-chloro-3-indoylphosphate-p-toluidin (BCIP) in staining buffer).
Courtesy Dr. Chrisopher Grefen, UK
DHAR1 ( Dehydroascorbate Reductase 1) the protein is induced by jasmonic acid and oxidative chemical stresses and is a key component of the ascorbate recycling system. Involved in redox homeostasis under biotic and abiotic inducers. Localized in mitochondria. Synonymes: glutathione-dependent dehydroascorbate reductase 1, chloride intracellular channel homolog 1, CLIC homolog 1, glutathione-dependent dehydroascorbate reductase 1, AtDHAR1, GSH-dependent dehydroascorbate reductase 1, mtDHAR, AT1G19570.
Witzel et al. (2017). Temporal impact of the vascular wilt pathogen Verticillium dahliae on tomato root proteome. J Proteomics. 2017 Oct 3;169:215-224. doi: 10.1016/j.jprot.2017.04.008.
Wang et al. (2014). Proteomic analysis of salt-responsive proteins in the leaves of mangrove Kandelia candel during short-term stress. PLoS One. 2014 Jan 8;9(1):e83141. doi: 10.1371/journal.pone.0083141. eCollection 2014.
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