2 μg K63-linked Di-Ubiquitin run on 4-12% SDS-PAGE gel under reducing conditions

Recombinant K63-linked Di-Ubiquitin

Protein

Product Code: TUB-058

Verified Applications: Ubiquitin chains exhibit diversity in length, linkage type, and associated cellular functions. K63-linked Di-Ubiquitin serves as a valuable reagent in assays involving ubiquitin-binding proteins and as a substrate for ubiquitin-specific deubiquitylating enzymes (DUBs). Optimal enzyme concentrations should be empirically determined based on the specific assay context.

Activity: Fully hydrolyzed by the K63-specific AMSH* deubiquitylase

Purity: 95%

Concentration: 58 μM

For Research Use Only (RUO)

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Protein Name: K63-linked Di-Ubiquitin

UniProt #: Ub

Predicted Molecular Weight: 17 kDa

Background and Additional Info

Background: Comprised of two 76 amino acid ubiquitin units joined via an isopeptide bond at K63. High-resolution structures (PDB 2JF5) reveal a left-handed, helical arrangement that is highly extended, while solution NMR shows the dimer samples multiple open and semi-closed states that are selectively stabilized on ligand binding. Unlike K48 chains, K63 polyubiquitin does not target substrates to the proteasome. Instead, it nucleates scaffold complexes in NF-κB (TRAF6-NEMO), DNA-damage tolerance (Rad5-PCNA), receptor endocytosis and innate-immune RIG-I signaling.

Formulation: 10 mM HEPES, pH 7.6

Shipping: The product is shipped with dry ice. Upon receipt, store it immediately at the temperature recommended below.

Storage and Stability: Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Aliquot and store ≤ -20°C (stable for 24 months from date of receipt).

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References: Varadan, R., et al., (2004) J Biol Chem 279:7055-63. PMID 14645257

Yoshikawa, A., et al., (2009) FEBS Lett 583:3317-22. PMID 19766637

Liu, Z., et al., (2015) eLife 4:e05767. PMID 26090905

Cao, L., et al., (2022) Cell Death Discov 8:410. PMID 36202787

Alternate Names: Human Lys63-linked Di-ubiquitin (K63-Ub₂, Ub₂-K63)

Protein Sequence: MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQKESTLHLVLRLRGG