AOX | AOX positive control/quantitation standard

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Aplikacje: western blot (WB)
Zalecane rozcieńczenie: 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.
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Przechowywanie: 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.
Uwagi: 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
Powiązane produkty: 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 antibodies
Dodatkowe informacje: 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.

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.
Literatura: to be added when available, standard available in May 2015