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SouthBayBio
ITCH E3 Ligase TR-FRET Kit
Product Details | |
---|---|
Synonyms | AIF4; AIP4; NAPP1 |
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 UBE2L3 (100μl) 100x ITCH (100μl) 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 ITCH leads to an increase in fluorescence emission at 665nm (Em-Acceptor) and decrease at emission wavelength 620nm (Em-Donor). Click here for a Typical Lot-specific Product Datasheet from the Original Manufacturer Our product description may differ slightly from the original manufacturers product datasheet. |
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 |
South Bay Bio’s ITCH TR-FRET Ubiquitin Kit provides a fast and sensitive method monitoring ubiquitin conjugation onto ITCH 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 ITCH 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.
- Real-Time Homogenous TR-FRET Ubiquitin Conjugation and Deconjugation Assay Platform: I. Tomasic: Poster, EMBO Conference Ubiquitin and SUMO, Cavtat-Dubrovnik (2017)