Ubiquitin (pSer65)

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Species Reactivity: Human  

Source: synthetic  

Formula: 50 mM HEPES pH 7.5, 150 mM sodium chloride, 2 mM dithiothreitol, 10% glycerol, 2% DMSO  

Concentration: 1 mg/ml  

Molecular Weight: 8.645kDa  

Protein Sequence: Accession number: P62990.1. For full protein sequence information download the Certificate of Analysis pdf.  

Quality Control Protein Identification: Confirmed by mass spectrometry.  

Quality Control Activity: Synthetic ubiquitin phosphorylated on Ser65 (ubiquitin (pSer65)) activates Parkin E3 ligase mediated ubiquitylation: Full-length Parkin (2 µg; Cat# 63-0048-025) was incubated at 30 °C with the ubiquitylation assay components Ube1 (0.1 µM; Cat# 61-0001) and Ube2L3 (1 µM; Cat# 62-0042) in the presence of 50 µM ubiquitin (comprising 20 μg of FLAG -ubiquitin mixed with nothing (lanes 1 and 2) or 5 μg of either enzymatically made ubiquitin (pSer65) (lanes 3 and 4), ubiquitin (lanes 5 and 6), synthetically made ubiquitin (pSer65) (Cat# 60-0202-050) (lanes 7 and 8) synthetically made ubiquitin (Cat# 60-0200-050) (lanes 9 and 10). Reactions were terminated after 60min by the addition of Lithium Dodecyl Sulfate (LDS) loading buffer and products were analysed by Sodium Dodecyl Sulfate (SDS) PAGE. Ubiquitin was detected using an anti-FLAG antibody.