Recombinant Human UbcH7/UBE2L3 - Ubiquitin Charged Protein
Recombinant Human UbcH7/UBE2L3 - Ubiquitin Charged Protein Summary
Product Specifications
Pre-charged UBE2L3 may be added directly to in vitro reactions containing E3 Ubiquitin ligases and ligase substrates (if applicable)—no ATP, E1 enzyme, or extra Ubiquitin are required. Reaction conditions will need to be optimized for each specific application. Note: Reducing agents including dithiothreitol (DTT) or mercaptoethanol ( beta ME) may cause unintended thiolytic release of Ubiquitin from the complex—care must be taken if these compounds are present in buffers. We suggest using a thioester friendly reductant such as TCEP if possible.
Accession # P68036, P0CG47
26 kDa (18 kDa UBE2L3, 8.6 kDa Ubiquitin)
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
E2-803
| Formulation | Approximately 0.45 mg/ml (25 μM) with respect to UBE2L3. Formulated in 50 mM HEPES pH 7.5, 50 mM NaCl, 1 mM TCEP |
| Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
Background: UbcH7/UBE2L3
Ubiquitin-conjugating Enzyme H7 (UbcH7), also known as Ubiquitin-conjugating Enzyme E2L 3 (UBE2L3), is a member of the Ubiquitinconjugating (E2) enzyme family. It has a predicted molecular weight of approximately 18 kDa. The human UbcH7 protein shares 100% amino acid (aa) sequence identity with the mouse and rat orthologs. UbcH7 has an E2 catalytic core domain that contains an active site cysteine residue and comprises 152 of its 154 aa residues. UbcH7 is catalytically active with HECT and RBR domaincontaining families of Ubiquitin ligases (E3s). UbcH7 localizes to both the nucleus and cytoplasm in human cells. In mice, its ortholog is expressed in many tissues including brain, muscle, heart, lung, lymph node, spleen, thymus, and testis. UbcH7 depletion results in an extended S phase and a reduced rate of proliferation, suggesting that it may play a role in the cell cycle. In humans, single nucleotide polymorphisms in UbcH7 are associated with systemic lupus erythematosus and Crohn's disease, suggesting that UbcH7 is important for proper immune system function.
This product is an enzymatically generated UbcH7/UBE2L3-Ubiquitin thioester complex that has been purified to remove E1 Ubiquitin Activating enzyme, uncharged UbcH7/UBE2L3, free Ubiquitin, and Mg2+-ATP. The product provides a convenient starting material for use in single-turnover "Ubiquitin Discharge Assays," eliminating the need to either inhibit the E1 Ubiquitin Activating enzyme with potentially confounding chemical treatments or remove ATP via enzyme additions.
- Das, R. et al. (2013) EMBO J. 32: 2504
- Koyano, F. et al. (2014) Nature 510: 162
- Liew, C.W. et al. (2010) Biochem. J. 431: 23
- Petroski, M.D. & Deshaies, R.J. (2005) Cell 123: 1107
- Saha, A. et al. (2011) Mol. Cell 42: 75
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