Specifications and Use
Fluorescent Substrate: Acetyl-dArg(3)-dGlu(3)-hexaminoyl-
K(Dabcyl)-PRYEAYKMGK(5FAM)-NH2
Catalog Number: PEPDAB063Am01, 1mg, 150 USD
Catalog Number: PEPDAB063Am05 5mg, 500 USD
Use:
This ADAM10 substrate is extremely selective and is used to assess activity only of ADAM10 No other ADAM or MMP metalloproteinases process this substrate. It is also the most sensitive ADAM10 substrate we have in addition to PEPMCA001, which is the MCA/DNP version. The ADAM10 substrate is based on the TENtide sequence discussed in “Active site determinants of substrate recognition by the metalloproteinases TACE and ADAM10. Biochemical Journal, Cristina I. Caescu, Grace R. Jeschke, and Benjamin E. Turk (2009), 424(1), 79-88”. However, we have added a charged tail in order to prevent the substrate from entering into the cell, thereby measuring only cell surface ADAM10. For information concerning its substrate specificity profile, see also our Product Sheets. Typically, the peptideADAM10 substrate 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 wells. For use with ADAM 10, the buffer should consist of 25mM Tris, pH 8, 6 x 10-4 Brij detergent, and 10mM CaCl2. Excitation and emission wavelengths are 485 and 530 nm respectively.
Molecular Weight:
2990.3 g/mol
Purity:
Greater than 95% as assessed by HPLC and Mass Spectrometry.
Solubility:
1 mg/ml in water with 10% Formic acid
Appearance:
Orange lyophilized powder
Counter Ion:
Trifluoroacetate
Shipping:
The peptide powder is shipped at room temperature.
Storage:
Upon receiving, the peptide should be stored at -20 oC or lower. 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 -20oC.
References:
Active site determinants of substrate recognition by the metalloproteinases TACE and ADAM10. Biochemical Journal, Cristina I. Caescu, Grace R. Jeschke, and Benjamin E. Turk (2009), 424(1), 79-88.