Ac-Pro-Ala-Leu-AMC (Ac-PAL-AMC)

Ac-PAL-AMC is a fluorogenic substrate of the branched amino acid preferring activity of the proteasome. It is preferentially cleaved by immunoproteasomes compared to constitutive proteasomes. The AMC fluorescence can be detected by a fluorimeter or a plate reader using excitation/emission wavelengths at 360 nm/460 nm, respectively.


This substrate can be used to determine the branched amino acid preferring activity of immunoproteasomes. Working concentration is 50 - 200 µM.

Additional Information

Product Name: Ac-Pro-Ala-Leu-AMC (Ac-PAL-AMC)
Also Known As: Ac-PAL-AMC
Catalog No.: G2111
Size: 25 mg
Molecular Weight: 498.6
Species: N/A
Source: Synthetic
Stock: Powder
Concentration: N/A
Quality Assurance: >95% by HPLC
Storage: Eligible for room temperature shipping. Store at -20°C upon receiving; avoid multiple freeze-thaw cycles after dissolving in DMSO.
PDF Data Sheet: Download PDF datasheet, MSDS
NCBI RefSeq: N/A
Image(s): G2110 G2111 image

20 nM constitutive (catalog # A1400) or immuno (catalog # A1500) bovine 20S proteasome was incubated with 120 nM PA28beta (catalog # A2200) for 15 min in 20 mM Tris, pH 7.1 at 37°C, 50 mM NaCl, 2 mM bME . Each substrate was prepared in the same buffer at 100 µM. Then 50 µl constitutive or immune 20S proteasome was mixed with 50 µl each of the substrates into a well of a 96-well plate, and AMC fluorescence was recorded immediately in a 20 min kinetic mode using the exciting/emission filter set at 360/460 nm, respectively. Linear slop of each curve was used to represent the 20S proteasome activity. Values from substrate alone were substracted as background. Error bars represent S.D. of three assays.
Shipping Method: Room temperature shipping
References: Blackburn, C., et al. (2010) Biochem. J. 430, 461.

Details

Ac-PAL-AMC is a fluorogenic substrate of the branched amino acid preferring activity of the proteasome. It is preferentially cleaved by immunoproteasomes compared to constitutive proteasomes. The AMC fluorescence can be detected by a fluorimeter or a plate reader using excitation/emission wavelengths at 360 nm/460 nm, respectively.
This substrate can be used to determine the branched amino acid preferring activity of immunoproteasomes. Working concentration is 50 - 200 µM.