Key features and details
- Assay type: Semi-quantitative
- Detection method: Fluorescent
- Platform: Microplate reader
- Assay time: 30 min
- Sample type: Cell culture extracts, Cell culture supernatant, Tissue Extracts
- Sensitivity: 40 ng/well
Product nameLysyl Oxidase Activity Assay Kit (Fluorometric)
Sample typeCell culture supernatant, Cell culture extracts, Tissue Extracts
Assay time0h 30m
Lysyl Oxidase Activity Assay Kit / LOX Activity Assay Kit (Fluorometric) (ab112139) provides a simple method to measure lysyl oxidase (LOX) activity in cell and tissue extracts from mammals and other species, as well as physiological solutions.
The LOX activity assay protocol uses a proprietary LOX substrate that releases hydrogen peroxide upon transformation by the LOX present in the sample. Hydrogen peroxide is in turn detected using a red fluorescence substrate for HRP-coupled reactions. This leads to increase in fluorescence that can be easily detected at Ex/Em = 540/590 nm in a fluorescence microplate reader.
This assay is semi-quantitative as it does not contain a LOX standard for calibration. When a known concentration of LOX is used, the assay can detect activity from as low as 40 ng of lysyl oxidase in solution.
The assay is highly sensitive and its unique detection method eliminates the interference that occurs in certain biological samples.
Lysyl oxidase (protein-lysine-6-oxidase, LOX, EC 220.127.116.11) is an extracellular copper-dependent enzyme that catalyzes formation of aldehydes from lysine residues in collagen and elastin precursors.
The activity of Lysyl oxidase in biological samples is traditionally assessed by tritium release end-point assays using radio isotope labeled collagen or elastin substrates.
Storage instructionsStore at -20°C. Please refer to protocols.
Components 500 tests Assay Buffer 1 x 50ml DMSO 1 x 200µl Horseradish Peroxidase 1 vial HRP Substrate 1 vial
Relative LOX activity levels in mouse tissue. Tissue samples were prepared following assay protocol. Protein concentration was determined and samples were diluted 2-30 fold. LOX activity levels were measured after 15 minutes incubation in a fluorometric plate reader at Ex/Em = 535/587 nm. LOX activity levels are relative to background noise (blank control).
LOX activity fluorescence (RFU) values vs quantity (µg). Recombinant human LOX2 was serially diluted 3.35-0.04 (1/3) and activity was measured following assay procedure.
LOX activity absorbance (OD) values vs quantity (µg). Recombinant human LOX2 was serially diluted 3.35-0.04 (1/3) and activity was measured following assay procedure. Colorimetric detection is approximately 10-times lower than fluorometric.
Typical lysyl oxidase (LOX) dose response curve. Known amounts of LOX were added to wells and reaction was run following assay protocol. Fluorescence was measured on a solid black 96-well plate using a Gemini fluiorescence microplate reader (Molecular Devices). Activity from as low as 40 ng/mL of LOX can be detected after 30 minutes incubation.
ab112139 has been referenced in 23 publications.
- Gudekar N et al. Metallothioneins regulate ATP7A trafficking and control cell viability during copper deficiency and excess. Sci Rep 10:7856 (2020). PubMed: 32398691
- Watanabe K et al. Group V secreted phospholipase A2 plays a protective role against aortic dissection. J Biol Chem 295:10092-10111 (2020). PubMed: 32482892
- Mahmutovic Persson I et al. Longitudinal Imaging Using PET/CT with Collagen-I PET-Tracer and MRI for Assessment of Fibrotic and Inflammatory Lesions in a Rat Lung Injury Model. J Clin Med 9:N/A (2020). PubMed: 33218212
- Bertero T et al. Tumor-Stroma Mechanics Coordinate Amino Acid Availability to Sustain Tumor Growth and Malignancy. Cell Metab 29:124-140.e10 (2019). PubMed: 30293773
- Rosell-Garcia T & Rodriguez-Pascual F Enhancement of collagen deposition and cross-linking by coupling lysyl oxidase with bone morphogenetic protein-1 and its application in tissue engineering. Sci Rep 8:10780 (2018). PubMed: 30018337