Additional Information
| Product Name: | Mca-KKVAPYPME-Dap(Dnp)-NH2 |
|---|---|
| Also Known As: | LFP |
| Catalog No.: | G1001 |
| Size: | 10 mg |
| Molecular Weight: | 1.53 KDa |
| Species: | N/A |
| Source: | Synthetic |
| Stock: | Powder |
| Concentration: | N/A |
| Quality Assurance: | >97% by HPLC |
| Storage: | Eligible for room temperature shipping. Store at -20°C upon receiving; avoid multiple freeze-thaw cycles after dissolving in DMSO. Protect from light. |
| PDF Data Sheet: | Download PDF datasheet, MSDS |
| NCBI RefSeq: | N/A |
| Image(s): | G1000 G1001 image (A) 4 nM constitutive 20S proteasome (catalog # A1400), 4 nM bovine 26S proteasome (catalog # A1200) or 4 nM bovine 20S proteasome + 32 nM PA700 (catalot # A1300) was incubated with 20 uM Mca-KKVAPYPME-Dap(Dnp)-NH2. The Mca fluorescence was recorded immediately in a 30 min kinetic mode using the exciting/emission filter set at 320/405 nm, respectively. See PDF Datasheet for details. (B) The slope values of curves in (A) that represent the activity of each sample were plotted as a bar graph. (C) 5 µg HEK293 cell lysates were incubated with or without 100 µM MG132 for 10 min at 37 0C, then mixed with 20 µM Mca-KKVAPYPME-Dap(Dnp)-NH2 to monitor the Mca fluorescence due to cleavage of the substrate. |
| Shipping Method: | Room temperature shipping |
| References: | Smith DM., et al, Mol. Cell, 2007, 27, 731. |
Details
Mca-KKVAPYPME-Dap(Dnp)–NH2 is a fluorogenic peptide substrate of proteasomes, in which the Mca fluorescence is internally quenched by Dap(Dnp). This substrate can be cleaved by mammalian, yeast or archaea activated proteasomes. Upon cleavage, the Mca fluorescence can be measured at excitation/emission wavelengths of 320/405 nm using a plate reader or fluorometer. Due to its long sequence as a nanomer, latent 20S proteasome has very weak activity on hydrolysis of Mca-KKVAPYPME-Dap(Dnp)–NH2, but it can be cleaved by activated proteasomes, including the 26S proteasome.
Mca-KKVAPYPME-Dap(Dnp)–NH2 can be used for 1) in vitro assaying 20S proteasome activation (gate opening) by assembling with a regulatory particle; and 2) determining the change of active proteasome activity in cells or tissues.
The working concentration is 10-20 µM.
