商品编号


J4120



品牌


UBPBio



公司


Ubiquitin-Proteasome Biotechnologies, LLC



公司分类


Proteasomes / Activators / Subunits



Size

1 Kit

商品信息

Proteasome Activity Fluorometric Assay Kit II is designed for assaying the proteasome’s chymotrypsin-like, trypsin-like and caspase-like activities in vitro using purified proteasome, cleared cell lysates or tissue extracts. The proteasome cleaves the supplied fluorogenic substrates and the released AMC fluorescence can be monitored using a plate reader or fluorometer at the excitation/emission wavelength of 360nm/460nm, respectively.

Additional Information






Product Name:

Proteasome Activity Fluorometric Assay Kit II



Also Known As:

N/A



Catalog No.:

J4120



Size:

1 Kit



Molecular Weight:

N/A



Species:

N/A



Source:

N/A



Stock:

20 mM Tris, 150 mM NaCl, 2 mM βME, 10% Glycerol



Concentration:

See tube label



Quality Assurance:

Enzyme >95% by SDS-PAGE; Inhibitor and substrate > 98% by HPLC and NMR



Storage:

Store at -80°C; avoid multiple freeze-thaw cycles



PDF Data Sheet:

PDF datasheet
,
MSD
S



NCBI RefSeq:

N/A



Image(s):

No



Shipping Method:

Dry ice shipping



References:

N/A




Details

* Each supplied fluorogenic substrate is sufficient for 165 x 150 ?l reactions. The supplied 20S proteasome is sufficient for 10 X 150 ?l reactions.

Description
Proteasome Activity Fluorometric Assay Kit II is designed for assaying the proteasome’s chymotrypsin-like, trypsin-like and caspase-like activities in vitro using purified proteasome, cleared cell lysates or tissue extracts. The proteasome cleaves the supplied fluorogenic substrates 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.
Suc-LLVY-AMC, Boc-LRR-AMC and Z-LLE-AMC are used to monitor the chymotrypsin-like, trypsin-like and caspase-like activities of the proteasome, respectively.
AMC is used to produce a standard curve using a concentration range of 10-100 pmol.
Dissolve 1 mg MG132 in 105 ?l DMSO to make a 20 mM stock solution before use, using 50-200 ?M MG132 to inhibit the proteasome in vitro.
The bovine 20S proteasome can be used as a positive control. A typical 150 ?l reaction contains 10 nM 20S proteasome, 50 ?M fluorogenic substrate in the assay buffer.

Components
? AMC (0.1 mM in DMSO)....................................................30 ?l ? 1000X SUC-LLVY-AMC (50 mM in DMSO)........................25 ?l
? 1000X BOC-LRR-AMC (50 mM in DMSO)........................25 ?l ? 1000X Z-LLE-AMC (50 mM in DMSO)...............................25 ?l
? 40X Bovine 20S Proteasome (400 nM).............................40 ?l ? MG-132..............................................................................1 mg
? DMSO..............................................................................200 ?l ? 20X Proteasome Assay Buffer........................................3x 1.35 ml