Specifications and Use
Fluorescent Substrate: Dabcyl-EHADLLAVVAK(5FAM)-NH2
Catalog Number: PEPDAB014m001, 1mg, 125 USD
Catalog Number: PEPDAB014m005, 5mg, 500 USD
Use:
This selective ADAM17 substrate is used to assess activity of ADAM17 using in vitro enzymatic and cell based assays. This ADAM17 substrate is based on the cleavage sequence in precursor TGF-alpha and is described by us in publication 1 below, ” Fluorescent substrates useful as high-throughput screening tools for ADAM9″. See our Product Sheets for substrate specificity profiles. Typically, the ADAM17 substrate is dissolved in DMSO to make a stock solution of about 10uM concentration. When used for in vitro assays, the ADAM17 substrate is often used at about 10mM 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 wells. For use with ADAM17, the buffer should consist of 25mM Tris, pH 8 and 6 x 10-4 Brij-35 detergent. Excitation and emission wavelengths are 485 and 530 nm respectively.
Molecular Weight:
1845.0 g/mol
Purity:
Greater than 95% as assessed by HPLC and Mass Spectrometry.
Solubility:
1 mg/ml in formic acid
Appearance:
Red lyophilized powder
Shipping:
The peptide powder is shipped at room temperature.
Storage:
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.
Stability:
Samples are sensitive to light and stable up to 6 months at -70oC.
References:
1.Fluorescent substrates useful as high-throughput screening tools for ADAM9.Marcia L Moss, Fred H Rasmussen, Raphael Nudelman, Peter J Dempsey, Jason Williams .Combinatorial chemistry & high throughput screening . 12/2009; 13(4):358-65. DOI:10.2174/138620710791054259
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.