AOX | AOX positive control/quantitation standard

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.Source of AOX standard: AOX standard source is Sphingomonas wittichii strain RW1, overexpressed in E.coli bearing an N-terminal his6 tag.

Standard curve: 3 loads are recommended (0.5, 2 and 4μl) .For most applications a sample load of 0.2 μg of chlorophyll/well will give a RbcL signal in this range.Positive control: a 2 μl load per well is optimal for most chemiluminescent detection systems. Higher standard concentration needs to be used to allow detection by Coomasie stains. Such gels with higher standard concentration can not be used for quantitation using chemiluminescence.This standard is stabilized and ready and does not require heating before loading on the gel. Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized. Allow the product several minutes to solubilize after adding water. Mix thoroughly but gently Take extra care to mix thoroughly before each use, as the proteins tend to settle with the more dense layer after freezing.

For reconstitution add 100 µl of milliQ water. Please note that this product contains 10% glycerol and might appear as liquid but is provided lyophilized.

The AOX calibrated protein standard can be used in combination with Agrisera global anti-AOX antibodies (AS04 054) to quantitate AOX from a wide range of species. Global antibodies are raised against highly conserved amino acid sequence. Quantitative western blot: detailed method description, video tutorial

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to 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.