Di-ubiquitin (K63-linked)

As indicated on product label or CoA when stored as recommended. Stable for at least 12 months after receipt when stored at -20°C. It is recommended that the product be kept at a moderately high protein concentration (>1mg/mL) and in the presence of buffer. If a more dilute reagent is required, the inclusion of an inert carrier protein (e.g. ovalbumin at 0.5-1.0mg/mL) in the storage buffer is recommended in order to minimise loss of the reagent due to absorption.

K 63 -linked di-ubiquitin (Ub 2) is made by incubating E1 and Ubc13-Mms2 with equal concentrations of ubiquitin capped at the future proximal chain terminus (Asp 77 -ubiquitin) and ubiquitin capped at the future distal chain terminus (K 48 R/K 63 R-ubiquitin) . The distal terminus of the resulting dimer is de-blocked by treatment with yeast ubiquitin hydrolase-1 (YUH-1) followed by purification by anion exchange chromatography.

For use in deubiquitinylating enzyme assays and polyubiquitin binding studies. Suggested uses: Deubiquitinylating enzyme substrates (general/linkage specific) . Investigation of polyubiquitin chain recognition by and interaction with ubiquitin binding proteins.