LOX | Lipoxygenase
AS06 128 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A.thaliana, G. max, L. longiflorum, L. luteus, O. europaea, O. sativa
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54 (subunit), 108 (native enzyme)
Glycine max, Lathyrus undulatus, Malus x domestica, Solanum tuberosum, Vicia fabaSpecies of your interest not listed? Contact us
Samples of Arabidopsis thaliana (2), Olea europaea (3), Lilium Longiflorum (4), Lupinus luteus (5) were ground in liquid nitrogen to a very fine powder using a mortar and pestle and resuspended in 1.5 ml of extraction buffer (4% SDS, 2% 2-mercaptoethanol, 2 mM PMSF, 100 mM Tris-HCl pH 8.5). The samples were incubated for 3 min at 80°C. Protein suspensions were clarified by centrifugation at 13,500 g for 10 min at room temperature and the resulting supernatants were used. Total proteins (25 µg per sample) were separated by SDS-PAGE on CriterionTMTGXTM Precast Gel (Bio-Rad, USA) using CriterionTM Cell apparatus (Bio-Rad). Proteins were electroblotted onto a PVDF membrane using Trans-Blot® TurboTM Transfer Pack (Bio-Rad) in a Trans-Blot® TurboTM Transfer System (Bio-Rad). The membrane was blocked for 1 h in solution containing 1 % (w/v) non-fat dry milk in TRIS-buffered saline (TBS) buffer, pH 7.4. The membrane was incubated in the primary antibody at a dilution of 1: 1000 in TBS buffer containing 1 % (w/v) non-fat dry milk over night at 4°C with agitation. A DyLight 488 conjugated anti-rabbit IgG (AS10 831, Agrisera), diluted 1:2000 in TBS buffer for 2 h, served as the secondary antibody. The signal was detected in a Pharos FX molecular imager (Bio-Rad). Line 1 contains LOX protein from Sigma.
Courtesy of Dr. Agnieszka Zienkiewicz, CSIC, Spain
Lipoxygenases (LOXs; EC 22.214.171.124, synonym: lipoxydases) are a family of enzymes that catalyze oxygenation of polyunsaturated fatty acids (PUFAs) into lipidhydroperoxides (LOOHs) involved in responses to stresses. LOXs has been found to play a role in plant growth and development, senescence as well as can be activated in response to environamental stress (drought, heavy metals). Synonymes: linoleate, oxygen oxidoreductase.
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