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SouthBayBio
Parkin E3 Ligase TR-FRET Kit
1250
CHF
CHF 1'250.00
In stock
SBB-KF0036-KI011 KitCHF 1'250.00
Figure 1: % Signal to Background of Continuous Real-Time TR-FRET Parkin titration (autoubiquitination): Serial dilutions of Parkin W403A from 50nM to 3.125nM and 300nM wt Parkin were mixed with UBA1, UBE2D3 and trf-Ub (pS65) mix. Reactions were initiated with addition of Mg-ATP.
Figure 2: Estimated Z-primes of Continuous Real-Time TR-FRET Parkin titration (autoubiquitination): Serial dilutions of Parkin from 50 nM to 3.125 nM mixed with UBA1, UBE2D3 and trf-Ub (pS65) mix. Reaction were initiated with addition of Mg-ATP.
| Product Details | |
|---|---|
| Synonyms | E3 Ubiquitin-protein Ligase Parkin; PARK2; Parkinson Juvenile Disease Protein 2; AR-JP; E3 Ubiquitin Ligase; LPRS2; Parkinson Disease Protein 2; PDJ; PRKN |
| Product Type | Kit |
| Properties | |
| Application Set | Compound Screening |
| Specificity |
Readout: Endpoint/Kinetic |
| Crossreactivity | Human |
| Quantity |
Sufficient Reagents for: |
| Kit Contains |
100x UBA1 (100μl) |
| 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 Parkin leads to an increase in fluorescence emission at 665nm (Em-Acceptor) and decrease at emission wavelength 620nm (Em-Donor). |
| 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 Parkin TR-FRET Ubiquitin Kit provides a fast and sensitive method monitoring ubiquitin conjugation onto both wild-type Parkin and a more active mutant W403A 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 Parkin 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
- 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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