Materials and Reagents

1. Cultured cells such as the macrophage line THP1
   
2. SensoLyte 520 TACE (α-Secretase) Activity Assay Kit *Fluorimetric* (AnaSpec, catalog number: 72085 )
   Components used from the kit:
   1. Component A QXL™ 520/5-FAM (TACE substrate, Ex/Em = 490 nm/520 nm)
      
   2. Component B 5-FAM, fluorescence reference standard (Ex/Em = 490 nm/520 nm)
      
   3. Component C assay buffer
      
3. Triton-X 100 (Sigma-Aldrich, catalog number: T8532 )
   
4. Phosphate buffered saline (PBS) (see Recipes)
   

Equipment

1. 96 well black microplate (for fluorescence analysis) (Greiner Bio-One GmbH, catalog number: 655077 )
   
2. Tube/vial unidirectional rotator (Glas-Col, catalog number: 099A RD4512 )
   
3. Cell lifter (polyethylene, sterile) (Corning Incorporated Costar, catalog number: 3008 )
   
4. 1.5 ml microcentrifuge tubes (polypropylene) (USA Scientific, catalog number: 1615-5599 )
   
5. Microplate fluorometer with detection range (excitation 490 nm and emission 520 nm) (settings recommended by SensoLyte 520 TACE Activity Assay Kit) (Thermo Fisher Scientific, Fluoroskan Ascent^TM) (used for TACE activity assay)

Procedure