Lysyl Oxidase Homolog 2 (LOXL2) Enzymatic LeadHunter Assay - TW

Procedure Summary:

Human recombinant LOXL2 enzyme expressed in mouse myeloma cells is used. Test compound and/or vehicle is preincubated with 4 µg/ml active enzyme in modified sodium borate buffer pH 8.0 for 15 minutes at 37°C. The reaction is initiated by addition of 300 uM cadaverine substrate and Amplex Ultrared for another 30 minute incubation period. Determination of the amount of resorufin formed is read spectrofluorimetrically at 544 nm/590 nm. Compounds are screened at 10 uM.

Substrate:

300 µM Cadavertine

Incubation:

30 min at 37°C

Control Inhibitor:

PXS-5153A

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 uM

Test Sample Requirements:

Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).

Minimum Order Quantity:

2