Plant/Algal cell antibodies / Environmental stress / Oxidative stress

product information

Alternative oxidases (AOX) are quinol oxidases located in the inner mitochondrial membrane of plants. They function as terminal oxidases in the alternate electron transport pathway, oxidizing ubiquinone to reduce oxygen to water.

alternative oxidase purified from Sauromatum guttatum (the voodoo lily)

store at -20°C; for long term storage at store at -80°C ; avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

western blot (WB), immunoprecipitation (IP), antibody column

AS04 054 | AOX1/2 | plant alternative oxidase 1 and 2 (polyclonal antibody for higher plants)

AS05 152 | AOX1 | alternative oxidase 1 (suitable for Chlamydomonas reinhardtii)

Recommended secondary antibody (for western blot with ECL detection)

antibody is supplied in cell culture medium with 1mM sodium azide as preservative

Protocol for plant mitochondria isolation can be found here.

application information

1 : 100 with standard ECL (WB, IP)

36-40 | 36-40 for Arabidopsis thaliana

Arabidopsis thaliana, Glycine max (AOX1,2,3), Sauromatum guttatum

dicots including: Nicotiana tabacum, monocots including: Oryza sativa, diatoms, fungus Aspergillus sp., Neurospora crassa, yeast, protozoa: Trypanosoma sp.

no confirmed exceptions from predicted reactivity known in the moment

Antibody is recognizing native and denatured AOX

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 extracts were prepared as described by Martinez-Garcia et al. (Plant J., 1999, 20:251-7).
Courtesy Dr. Janusz Piechota, Wrocław University, Poland