Available ordering format
Lyophilized
Immunogen
to be determined
Raised in
N/A
Clonality
N/A
Clone
N/A
Purification
Affinity purified
How to reconstitute
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.
Storage condition
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.
Verified applications
western blot (WB)
Connected products
Primary antibodies matching AOX standard are: AS04 054 | AOX1/2 | plant alternative oxidase 1 and 2, rabbit antibodycollection of other protein standardscollection of matching global antibodiesPlant and algal protein extraction buffer
Recommended dilutions for use
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.
Molecular weight (expected | аpparent)
27 kDa
Verified reactivity
to be determined
Possible reactivity
to be determined
No reactivity
to be determined
Supplementary information
Concentration: 0.1 pmol/µl. After re-constitution with sterile milliQ water, the final concentration of the AOX monomer is.0.1 pmol/µl. While a dimer is present in the lane, only the 27 kDa monomer contributes to the calibration.Protein standard buffer composition: Glycerol 10%, Tris Base 141 mM, Tris HCl 106 mM, LDS 2%, EDTA 0.51 mM, SERVA® Blue G250 0.22 mM, Phenol Red 0.175 mM, pH 8.5, 0.1mg/ml PefaBloc protease inhibitor (Roche), 50 mM DTT.This standard is ready-to-load and does not require any additions or heating. It needs to be fully thawed and thoroughly mixed prior to using. Avoid vigorous vortexing, as buffers contain detergent. Following mixing, briefly pulse in a microcentrifuge to collect material from cap.
References
to be added when available, standard available in May 2015
Scientific context
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.
Notes
The AOX calibrated protein standard can be used in combination with Agrisera global anti-AOX antibiodies (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
Protein number
Refer to NCBI
TAIR number
Refer to NCBI
Tissue
control
Description
Isotype or positive controls by peptides, antibodies and deactivated samples.Positive controls are the same as the target vector or antibody or protein and can be spiked to the sample before the analysis starts.
Group
positif