Ubiquitin Enzyme Assay

UC108: TR-FRET Based In Vitro E3 Ligase Customizable Kit

$995.00

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SKU: UC-0108 Categories: , Tags: ,

Specifications

The TR-FRET In vitro E3 activity kit has been developed to be simple, fast, and easy to use while being robust and sensitive. As E3 ligases autoubiquitinate they build long polyubiquitin chains; our unique assay will detect these chains as they are being built. This assay is homogenous and compatible for High-throughput Screening (HTS).

This product is protected by one or more US or Foreign patents. Please read the Limited Use Label License to learn more. By purchasing this product, the purchaser agrees to comply to these terms.

Kit Components

  •  10x Assay Buffer
  • Choice of E3
  • Choice of E2
  • Ubiquitin Activating Enzyme E1
  • Ubiquitin Mixture (20x)
  • ATP
  • TR-FRET Detection Mix
  • Positive Control E3 (Carp2 with UBE2D3)

Info

Storage All components should be stored at -80°C.
Stability All components, except for the pre-coated plates, are stable for at least 2 months when stored at -80°C.
Plates must be used within 30 days.

The selection of appropriate streptavidin reagent is critical to generating a robust signal. We do not recommend using avidin-horseradish peroxidase (HRP) conjugates.

Additional information

E2

UBE2D3 (UbcH5c), UBE2L3 (UbcH7), UBE2K (UbcH1), Ubc13, Mms2 (yeast), UBE2D2 (UbcH5b, E2-17K2), UBE2R1 (CDC34), UBE2R2 (Ubc3B), Non-tagged UBE2D1, His6-SUMO tagged UBE2D1, UBE2E2 (UbcH8), UBE2E3 (UbcH9), His6-tagged UBE2H (E2-20K), His6-SUMO tagged UBE2H (E2-20K), UBE2F (NCE2), UBE2M (Ubc12), UBE2N (Ubc13), Non-tagged UBE2T (PIG50), His6-SUMO tagged UBE2T (PIG50), UBE2V2 (MMS2, UEV-2), UBE2S (E2EPF), UBE2C (Ubc10), His6-tagged UBE2W (Ubc 16), UBE2Q2, His6-tagged UBE2A, Non-tagged UBE2G2 (Ubc7), His6-SUMO tagged UBE2G2 (Ubc7), His6-tagged UBE2I (Ubc9), UBE2L6 (UbcH8), UBE2Z (Use1)
E3

ITCH, Traf6, GP78, WWP2, MDM2, Hrd1, MuRF1, MuRF2, MuRF3, E6AP, Praja1, CARP2, Parkin

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Applications

  • Quantitative Analysis  of  Ubiquitin E2 Conjugation or E3 Ligase Activity
  • Demonstration of novel E2 or E3 Activity
  • High Throughput Screening (HTS) of antagonist or agonists of ligase activity
  • Monitor enzyme activity during purification
  • Monitor E3 activity in solution phase, with the E2 of your choice
  • Plate-based format is amenable to high-throughput screening (HTS)
  • Detection system provides robust readout for E3 ligase activity
  • E3LITE assay utilizes non-radioactive reporter substrates
  • E3LITE assay does not require excitation in the UV range (reducing false positive rate)

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