Specifications and Use
Fluorescent Substrate: Dabcyl-HGDQMAQKSK(5FAM)-NH2
Catalog Number: PEPDAB013m001, 1mg, 125 USD
Catalog Number: PEPDAB013m005, 5mg, 500 USD
This fluorescent ADAM8 substrate was originally described by us in the publication, ” Fluorescent substrates for the proteinases ADAM17, ADAM10, ADAM8 and ADAM12 useful for high throughput screening”. This ADAM substrate has been used to assess activity of ADAM8 using in vitro enzymatic and cell based assays (See publications 1-4) as it is a highly sensitive and selective substrate for this enzyme. (Please also see our product sheets or catalog, for the substrate specificity profile). Typically, the peptide is dissolved in DMSO to make a stock solution of about 10mM concentration. When used for in vitro assays, the substrate is often used at about 10uM concentration. Remember to keep the DMSO concentration in the final reaction at 1% or below, to avoid DMSO effects on the reaction, and remember to have an equivalent percentage of DMSO in the background well. For use with ADAM8, the buffer should consist of 50mM Tris, pH 8, 150mM NaCl, 10mM CaCl2 , and 6 x 10-4 Brij detergent. Excitation and emission wavelengths are 485 and 530 nm respectively.
Greater than 92% as assessed by HPLC and Mass Spectrometry.
1 mg/ml in water with 10% Formic acid
Red lyophilized powder
The peptide powder is shipped at room temperature.
Upon receiving, the peptide should be stored at -70 oC. Avoid repeated freeze-thaw cycles. If dissolved in liquid (such as DMSO), aliquot into separate tubes to minimize the number of freeze-thaw cycles.
Samples are light sensitive and stable up to 6 months at -70oC.
1.Fluorescent substrates for the proteinases ADAM17, ADAM10, ADAM8, and ADAM12 useful for high-throughput inhibitor screening. Marcia L Moss, Fred H Rasmussen.(2007) Analytical Biochemistry;366(2):144-8. PMID 17548045
2.Proteolytic Activity Matrix Analysis (PrAMA) for simultaneous determination of multiple protease activities. Miles A Miller, Layla Barkal, Karen Jeng, Andreas Herrlich, Marcia Moss, Linda G Griffith, Douglas A Lauffenburger Integrative Biology 12/2010; 3(4):422-38.
3.Potential of Fluorescent Metalloproteinase Substrates for Cancer Detection. Roopali Roy,1,5 David Zurakowski,2,5 Susan Pories,3 Marcia L. Moss,4,* and Marsha A. Moses1,5Clin Biochem. 2011 December; 44(17-18): 1434–1439.
4.ADAM8 as a drug target in pancreatic cancer. Uwe Schlomann, Garrit Koller, Catharina Conrad, Taheera Ferdous, Panagiota Golfi, Adolfo Molejon Garcia, Sabrina Höfling, Maddy Parsons, Patricia Costa, Robin Soper, Maud Bossard, Thorsten Hagemann, Rozita Roshani, Norbert Sewald, Randal R. Ketchem, Marcia L. Moss, Fred H. Rasmussen, Miles A. Miller, Douglas A. Lauffenburger, David A. Tuveson, Christopher Nimsky & Jörg W. Bartsch Nature Communications, 6,Article number: 6175, doi:10.1038/ncomms7175