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application information | ||
recommended dilution | 1 : 100 with standard ECL (WB, IP) | |
expected | apparent MW | 36-40 | 36-40 for Arabidopsis thaliana | |
confirmed reactivity | Arabidopsis thaliana, Glycine max (AOX1,2,3), Sauromatum guttatum | |
predicted reactivity | dicots including: Nicotiana tabacum, monocots including: Oryza sativa, diatoms, fungus Aspergillus sp., Neurospora crassa, yeast, protozoa: Trypanosoma sp. | |
not reactive in | no confirmed exceptions from predicted reactivity known in the moment | |
additional information | Antibody is recognizing native and denatured AOX | |
selected references | Kakizaki and Ito (2013). Engineering Plant Alternative Oxidase Function in Mammalian Cells: Substitution of the Motif-like Sequence ENV for QDT Diminishes Catalytic Activity of Arum concinnatum AOX1a Expressed in HeLa Cells. Costa-de-Oliveira et al. (2012). An alternative respiratory pathway on Candida krusei: implications on susceptibility profile and oxidative stress. FEMSyeat Res. January 23 ( ahead of print). Elthon et al. (1989). Monoclonal antibodies to the alternative oxidase of higher plant mitochondria. Plant Physiol. 89:1311-1317. |
application example 25 μg of Arabidopsis thaliana mitochondrial wild type fraction (1) mitochondrial fraction from a mutant with increased AOX level (2), total wild type leaf extract (3), total leaf extract from AOX overproducing mutant (4) were separated on 10% gel and blotted on nitrocellulose membrane using wet transfer (0.22% CAPS, pH 11). Filters where blocked (1.5h) in 5% milk in TBST (1X TBS, 0,1% Tween 20), incubated with 1: 100 anti-AOX monoclonal antibodies (2h in TBST) followed by 1 h incubation with 1: 10 000 Agrisera secondary anti-mouse HRP-coupled antibodies (AS09 627) and visualized with standard ECL on Kodak autoradiography film for 15-60 s. Mitochondria were isolated as described by Urantowka et al. (Plant Mol Biol, 2005, 59:239-52). Mitochondrial pellets were suspended in 1X Laemmli buffer (5% beta-mercaptoetanol, 3.7% glycerol, 1.1% SDS, 23 mM Tris- HCl pH 6.8, 0.01% bromophenol blue), heated (95°C, 5 min.) and centrifuged (13 000rpm, 1 min.). Leaf extrac 手機(jī)版:AOX | alternative oxidase
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