Deubiquitinase Activity Assay Kit (Fluorometric) (BN00721)

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BN00721
  • Deubiquitinase Activity Assay Kit (Fluorometric) (BN00721)
  • Deubiquitinase Activity Assay Kit (Fluorometric) (BN00721)
  • Deubiquitinase Activity Assay Kit (Fluorometric) (BN00721)
€613

Description

ELISA Kit Technical ManualMSDS

Deubiquitinase Activity Assay Kit (Fluorometric)

Cell activity and viability is tightly regulated by controlling the production and degradation of the thousands of different proteins in the cell. The proteasome is responsible for the majority of cellular protein degradation; however, drugs targeting the proteasome can have side effects caused by the lack of specificity associated with inhibiting the proteasome itself. Altering the ubiquitination state of target proteins is thus appealing as an alternative approach. Modification of the ubiquitin-mediated proteasome pathway has been shown to be a valid mechanism for treating a variety of diseases, all of which involve dysregulation of cellular proteostasis. As such, it is imperative that these ubiquitination signals also be reversible. The enzymes responsible for cleavage, and hence removal of ubiquitin from ubiquitinated proteins, are known as de-ubiquitinating enzymes, or DUBs. They are proteases that hydrolyze the isopeptide bond between an ubiquitin moiety and a lysine residue on its target protein. By removing the ubiquitin molecule, the protein escapes the fate of proteasomal degradation and remains a viable factor in the cell. Assay Genie's Deubiquitinase Activity Assay Kit provides a straight-forward and general measure of deubiquitinase activity by utilizing a fluorescent deubiquitinase substrate to detect activity as low as 0.25 µU with purified enzyme.

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Figure: (a) AMC Standard Curve; (b) Time course using positive control DUB as described; (c) Example of determination of DUB activity in tissue lysates. Rat tissue samples (10 mg each) were resuspended in DUB Assay Buffer with DTT (100 l), homogenized, and clarified by centrifugation. The DUB activities for Rat Kidney and Liver lysates, in mU/mg, were determined to be 0.76 and 3.31, respectively. Assays were performed following the kit protocol.

Key Information Description

Product SKU

BN00721

Size

100 assays

Detection Method

Fluorescence (Ex/Em 350/440 nm)

Species Reactivity

Mammalian

Applications

  • Measurement of DUB activity in various tissues/cell extracts
  • Determination of DUB activity associated with pathological conditions
  • Characterization of activity of purified DUB enzymes

Features and Benefits

  • Simple procedure; takes only ~40 min
  • Fast and convenient

Kit Components

  • DUB Assay Buffer
  • 1 M DTT
  • DUB Substrate (in DMSO)
  • DUB Positive Control
  • AMC Standard (1 mM)
  • White 96-well Half-Area Plate

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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