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SouthBayBio
TRF-Ubiquitin Mix (100X) (400 reactions)
1233
CHF
CHF 1'233.00
In stock
SBB-TR0051-4H-11 VialCHF 1'233.00
Figure 1: % Signal to Background of Continuous Real-Time TR-FRET E3 Ligase titration (autoubiquitination): An example using serial dilutions of GST-MDM2 from 200nM to 12.5nM mixed with UBA1, UBE2D3 and trf-Ub mix. Reaction was initiated with addition of Mg-ATP.
Figure 2: Estimated Z-primes of Continuous Real-Time TR-FRET E3 Ligase titration (autoubiquitination): Example of serial dilutions of GST-MDM2 from 200nm to 12.5nM mixed with UBA1, UBE2D3 and trf-Ub mix. Reaction was initiated with addition of Mg-ATP.
| Product Details | |
|---|---|
| Synonyms | TRF-Ub Mix |
| Product Type | Protein |
| Properties | |
| Source/Host | E. coli |
| Crossreactivity | Human |
| Application |
TR-FRET: 400 x 20μl reactions or 160 x 50μl reactions |
| MW | 8.5 to 8.9 kDa |
| Purity | ≥99% (LCMS) |
| Formulation | Liquid. In 50mM HEPES pH 7.5, 100mM NaCl. |
| Other Product Data |
This product is a mix of Europium-Cryptate Ubiquitin (SBB-TR0014), Cy5-Ubiquitin (SBB-TR0015), and wild-type Ubiquitin (SBB-UP0013) in a ratio optimized for TR-FRET based conjugation experiments where long polyubiquitin chains are formed. Under conditions where short ubiquitin chains are formed, a different mix may be required to optimize signal to background ratio. Enzymatic incorporation of the labeled ubiquitins into chains conjugated onto a substrate protein 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. Protect from light. |
| Use/Stability | Stable for at least 1 year after receipt when stored at -80°C. |
| Documents | |
| MSDS | Inquire |
| Product Specification Sheet | |
| Datasheet |
 Download PDF |
Description
Ubiquitin is a highly conserved protein that plays a major role in the ubiquitination pathway, which is conserved from yeast to mammals. Ubiquitination, the conjugation of ubiquitin to other proteins through a covalent bond between its C-terminal glycine and the ε-amino group of lysine residues or the α-amino group of an N- terminal methionine onto proteins is essential for many cellular processes primarily linked to protein degradation. This process involves three steps with specific groups of enzymes in an ATP depended manner, which are activation with ubiquitin-activating enzymes (E1s), conjugation with ubiquitin-conjugating enzymes (E2s) and ligation with ubiquitin ligases (E3s).
This product is a mix of Europium Cryptate Ubiquitin (SBB-TR0014), Cy5-Ubiquitin (SBB-TR0015), and wild-type Ubiquitin (SBBUP0013) in a ratio optimized for TR-FRET based conjugation experiments where long polyubiquitin chains are formed. Under conditions where short Ubiquitin chains are formed, a different mix may be required to optimize signal to background ratio. Enzymatic incorporation of the labeled ubiquitins into chains conjugated onto a substrate protein leads to an increase in fluorescence emission at 665nm (Em acceptor) and decrease at emission wavelength 620nm (Em donor).
Product References
- Imidodiphosphinate ligands as antenna units in luminescent lanthanide complexes. s. Magennis, et al.; Chemical Communications 1, 61 (1999)
- Ubiquitin Ligases: Structure, Function, and Regulation. N. Zheng & N. Shabek; Annual Review of Biochemistry (2017)
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