SouthBayBio

MDM2 E3 Ligase TR-FRET Kit

CHF 1'250.00
In stock
SBB-KF0030-KI011 KitCHF 1'250.00
More Information
Product Details
Synonyms HDM2
Product Type Kit
Properties
Application Set Compound Screening
Specificity Readout: Endpoint/Kinetic
Label or Dye: Europium-Cryptate & Cy5
Detection Method: TR-FRET
Substrate Properties: Protein-based Substrate. Typical experimental concentration is 1x.
Excitation/Emission (nm): 304/620 & 304/665
Crossreactivity Human
Quantity Sufficient Reagents for:
400 x 20μl reactions
160 x 50μl reactions
Kit Contains 100x UBA1 (100μl)
100x UBE2D3 (100μl)
50x GST-MDM2 (200μl)
100x TRF-Ubiquitin Mix (100μl)
10x Reaction Buffer (1ml)
10x Mg-ATP (1ml)
1 x 384-well white low-volume microplate
Other Product Data The kit uses ubiquitin labeled with either Europium-Cryptate or Cy5 as FRET pair donor and acceptor fluorophores respectively, completely eliminating the need for antibody based detection setups. Enzymatic incorporation of the labeled ubiquitins into chains conjugated onto MDM2 leads to an increase in fluorescence emission at 665nm (Em-Acceptor) and decrease at emission wavelength 620nm (Em-Donor).

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Declaration Manufactured by South Bay Bio.
Shipping and Handling
Shipping DRY ICE
Short Term Storage -20°C
Long Term Storage -80°C
Handling Advice Aliquot to avoid freeze/thaw cycles.
Avoid freeze/thaw cycles.
Documents
Manual No
MSDS Inquire
Product Specification Sheet
Datasheet Download PDF
Description

South Bay Bio’s MDM2 TR-FRET Ubiquitin Kit provides a fast and sensitive method monitoring ubiquitin conjugation onto GST-MDM2 in solution, resulting from an enzymatic ubiquitin cascade without the need of running and staining an SDS gel. The kit enables continuous TR-FRET detection of ubiquitin chain formation onto MDM2 in a real-time detection setup, or in an end-point configuration if desired. TR-FRET uses the extended fluorescence emission decay lifetimes typical of rare-earth lanthanides to impart a short time-delay between FRET donor excitation and emission. This delay provides a means to separate “true” signal from short-lived background fluorescence, and reduce interference from compound fluorescence and other assay artifacts.

Product References
  1. Real-Time Homogenous TR-FRET Ubiquitin Conjugation and Deconjugation Assay Platform: I. Tomasic: Poster, EMBO Conference Ubiquitin and SUMO, Cavtat-Dubrovnik (2017)
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