Lysyl Oxidase Activity Assay
The Assay Genie Lysyl Oxidase Activity Assay allows for quantitative evaluation of lysyl oxidase activity of purified enzyme and its inhibitors as well as secreted endogenous enzyme in cell media. Enzyme activity is detected upon oxidation of substrate, which results in release of reaction intermediates. Reaction intermediates are subsequently detected by Assay Genie's LOX Probe, resulting in fluorescent signal that can be measured at Ex/Em= 535/587 nm. Each kit includes a positive control as well as a specific lysyl oxidase inhibitor that allows the user to correct each sample for any non-specific signal. The Lysyl Oxidase Activity Assay can detect as little as 40 ng of recombinant enzyme in vitro.
Lysyl Oxidase Activity Assay Kit Data
|Product Name:||Lysyl Oxidase Activity Assay|
|Detection Method:||Fluorometric (Ex/Em = 535/587 nm)|
|Sensitivity:||40 ng of recombinant enzyme in vitro|
|Sample Type:||Purified protein, cell media.|
|Storage:||Store the kit at -20°C, protect from light|
|Shipping Conditions:||Gel Pack|
|Usage:||For research use only. Not for use in humans.|
Lysyl Oxidase Activity Assay Kit Components
|LOX Assay Buffer||45 ml|
|LOX Substrate||100 µl|
|LOX Developer||1 vial|
|LOX Probe (in DMSO)||100 µl|
|LOX Positive Control||1 vial|
|LOX Inhibitor||1 vial|
|LOX Standard (0.88 M)||100 µl|
Figure: a) Enzyme activity at varying concentrations of recombinant Lysyl Oxidase b) representative activity curve for cell media sample and cell media spike standard at 37°C, where cell media is test sample, cell media + inhibitor is negative control and 150 pmol spike standard is sample + spike standard.
Lysyl Oxidase Background
Lysyl oxidase is a cell-secreted amine oxidase that catalyzes the final step of collagen and elastin cross-linking. As a result, lysyl oxidase is required for the biosynthesis of functional extracellular matrices. Inhibition of lysyl oxidase can result in osteolathyrism, a condition characterized by fragile connective tissue and paralysis. Recent work has also suggested lysyl oxidase to play an important role in cancer development: Lysyl oxidase promotes tumor growth and progression in vivo, cancer cell invasion, and premetastatic niche formation.