ADAM10 Fluorogenic Assay Kit

• Background:

  ADAM10 (A disintegrin and a metalloprotease 10), also known as, is one of the 22 members of the ADAM family of proteins found in humans and part of the zinc protease superfamily. It has two main known functions, acting as a metalloprotease and in adhesion. This protein is synthetized as an inactive proform, which is processed in the Golgi by removal of its inhibitory domain. It can act on several substrates, one of them being the Notch receptor. After ligand binding to the Notch receptor, ADAM10 cleaves the ectodomain, leaving the membrane regions to be processed by γ-secretase and the release of a NICD (notch intracellular domain) . ADAM10 is itself regulated at multiple levels, such as the transcription level by SP1 (specificity protein 1) and a retinoic-acid -responsive element, amongst others. It is involved in cleavage of proteins that are near an extracellular membrane leaflet, and thus regulates the turnover of proteins involved in adhesion and signaling, such as cadherins and receptors. It is expressed in the brain, breast, liver, epithelia, and others. It has been implicated in neurodegeneration, cancer and immune disorders. APP (amyloid precursor protein) is a substrate for ADAM10, and upregulation of this metalloprotease can bring benefits to Alzheimer’s disease (AD) patients. ADAM10 also has VE-cadherin, TNFα (tumor necrosis factor α), IL-6R, CXCL16 (chemokine C-X-C motif ligand 16) and CX3CL1 as substrates, participating in immune responses and contributing to auto-immune disorders such as RA (rheumatoid arthritis), SLE (systemic lupus erythematosus) and others.  The development of therapies targeting this protein will no doubt result in advances in ADAM-10 related diseases.

• Description:

  The ADAM10 Fluorogenic Assay Kit is designed to measure ADAM10 (A disintegrin and a metalloprotease 10) protease activity for screening and profiling applications. The assay kit comes in a convenient 96-well format, with enough recombinant ADAM10, fluorogenic substrate and solution, and assay buffer for 100 enzyme reactions.Need us to run inhibitor screens or profile your compounds against ADAM10? Check out ourProtease Screening Services.Figure 1: Illustration of the mechanism behind the ADAM10 Fluorogenic Assay Kit.ADAM10 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.

• UniProt:

  O14672

• Applications:

  Study enzyme kinetics and screen small molecule inhibitors of ADAM10 for drug discovery and high throughput screening (HTS) applications.

• Format:

  Catalog #NameAmountStorageADAM10*3 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.