Proteasome Activity Fluorometric Assay Kit I

Proteasome Activity Fluorometric Assay Kit I was designed for assaying the constitutive proteasomes' chymotrypsin-like activity in vitro using purified proteasome, cell lysates or tissue extracts. The proteasome cleaves Suc-LLVY-AMC, and the released AMC fluorescence can be monitored using a plate reader or fluorometer at the excitation/emission wavelength of 360nm/460nm, respectively.


The supplied Suc-LLVY-AMC is sufficient for 250 X 100 µl reactions.

Additional Information

Product Name: Proteasome Activity Fluorometric Assay Kit I
Also Known As: N/A
Catalog No.: J4110
Size: 1 Kit
Molecular Weight: N/A
Species: N/A
Source: N/A
Stock: N/A
Concentration: See tube label
Quality Assurance: Inhibitor and substrate > 98% by HPLC and NMR; activity assay using cell lysates.
Storage: Store at -80°C; avoid multiple freeze-thaw cycles
PDF Data Sheet: PDF datasheet, protocol
NCBI RefSeq: N/A
Image(s): J4110 Image:
(Click image to enlarge)

AMC fluorescence recorded in reactions containing 50 µg HEK293T cell lysates, 50 µM Suc-LLVY-AMC in the presence or the absence of 100 µM MG132.

Shipping Method: Dry ice shipping
References: N/A

Details

The supplied Suc-LLVY-AMC is sufficient for 250 X 100 µl reactions.

Description

Proteasome Activity Fluorometric Assay Kit I is designed for assaying the proteasome’s chymotrypsin-like activity in vitro using purified proteasome, cleared cell lysates or tissue extracts. The proteasome cleaves Suc-LLVY-AMC and the released AMC fluorescence can be monitored using a plate reader or fluorometer at the excitation/emission wavelength of 360nm/460nm, respectively. The amount of the cell lysates or tissue extracts to be used in the reaction should be optimized.

AMC is used to produce a standard curve using a concentration range of 10-100 pmol.

Components

• AMC (0.1mM in DMSO)................................................30 µl
• 1000X SUC-LLVY-AMC (50 mM in DMSO)...................25 µl
• HEK293T cell lysates (5mg/ml).....................................100 µl
• MG-132..........................................................................1 mg
• MgCl2 (1M)....................................................................250 µl
• ATP (500 mM)................................................................100 µl
• DMSO............................................................................200 µl
• 20X Proteasome Assay Buffer.......................................1.35 ml