ADAM17 Fluorogenic Assay Kit

• Background:

  ADAM17 (A disintegrin and a metalloprotease 17), also known as TACE (TNF-α converting enzyme), is one of the 22 members of the ADAM family of proteins found in humans and part of the zinc protease superfamily. It is involved in development, immunity, inflammation and tumorigenesis. ADAM17 is found in the brain, heart, kidney and skeletal muscle, amongst others. It has more than 80 substrates, ranging from EGFR (epidermal growth factor receptor) ligands, TNF-α (tumor necrosis factor alpha) to adhesion molecules and amyloid precursor protein, and it is thus involved in multiple pathways. Its activity is highly regulated, with low activity resulting in issues with regeneration due to low EFGR signaling, while increased activity can lead to tumor development. This protein is formed in the ER (endoplasmic reticulum) as a proform and matures in the Golgi apparatus where the prodomain is cleaved by pro-protein convertases. Patients with RA (rheumatoid arthritis) have elevated activity of ADAM17 in the synovial tissue. Because of the roles it plays, targeting this protein in diseased tissues without affecting the healthy tissues is a major concern.  The development of therapies targeting this protein in specific tissues will no doubt result in advances in ADAM-17 related diseases.

• Description:

  The ADAM17 Fluorogenic Assay Kit is designed to measure ADAM17 (A disintegrin and a metalloprotease 17) protease activity for screening and profiling applications. The assay kit comes in a convenient 96-well format, with enough recombinant ADAM17, fluorogenic substrate and solution, and assay buffer for 100 enzyme reactions.Need us to run inhibitor screens or profile your compounds against ADAM17? Check out ourProtease Screening Services.Figure 1: Illustration of the mechanism behind the ADAM17 Fluorogenic Assay Kit.ADAM17 is incubated with a fluorogenic substrate which is an internally quenched fluorogenic substrate. Proteolysis releases the highly fluorescent Mca from the quencher. Fluorescence intensity increases proportionally to the activity of the protease.This product has beencited 2 times.

• UniProt:

  P78536

• Applications:

  Study enzyme kinetics and screen small molecule inhibitors of ADAM17 for drug discovery and high throughput screening (HTS) applications.

• Format:

  Catalog #ComponentAmountStorageADAM173 µg-80°CADAM Fluorogenic Substrate I50 µl-80°C780011X ADAM Assay Buffer5 ml-20°C79685Low binding, black 96-well plate1RoomTemp*The concentration of the protein is lot-specific and will be indicated on the tube.

• Shipping Conditions:

  -80°C

• Storage Conditions:

  This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.

• Notes:

  Troubleshooting GuideVisitbpsbioscience.com/assay-kits-faqfor detailed troubleshooting instructions. For all further questions, please email[email protected]

• Contraindications:

  The final concentration of DMSO in the assay should not exceed 1%.Fluorescent compounds that have λex=485 nm (5 nm bandwidth) and detection at λem=530 nm (5 nm bandwidth) can interfere with the readouts.The presence of strong acids or bases, ionic detergents and high salt should be avoided.