Ubiquitin-Rhodamine is prepared by the C-terminal derivatization of ubiquitin with modified rhodamine-110 and has been shown to be a useful and sensitive fluorogenic substrate for wide range of deubiquitinylating enzymes (DUBs), including ubiquitin C-terminal hydrolases (UCHs) and ubiquitin specific proteases (USPs). Ubiquitin-rhodamine is particularly useful for studying deubiquitinylating activity where detection sensitivity or continuous monitoring of activity is essential. The long excitation and emission wavelengths of rhodamine 110 (Excitation 485nm / Emission 520nm) avoid interference often caused at shorter wavelengths by small molecules and tissue culture components.
Product Details
Sequence: | Accession number: P62988; length: 76 amino acid residues. |
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MW: | ~8.9kDa |
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Source: | Produced in E. coli and covalently attached to modified rhodamine-110. |
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UniProt ID: | P0CG47 (UBB), P0CG48 (UBC), P62979 (RPS27A), P62987 (UBA52) |
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Formulation: | Liquid. In dimethylsulfoxide. |
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Purity: | ≥95% (HPLC) |
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Purity Detail: | Please note: considerable efforts are taken to ensure that ubiquitin-rhodamine is homogeneous and not contaminated with ubiquitin and ubiquitin deletions, which are common in material obtained from other sources. The chromatogram below demonstrates the homogeneity of Prod. No. BML-SE761. |
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Application Notes: | Can be used for the following applications: 1. Substrate for deubiquitinylating enzyme activity assays. 2. Identification/confirmation of enzyme deubiquitinylation activity. 3. Investigation of deconjugating enzyme substrate specificity in comparison with alternative UBL substrates. |
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Shipping: | Shipped on Dry Ice |
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Short Term Storage: | -80°C |
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Long Term Storage: | -80°C |
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Use/Stability: | Stable for at least one year after receipt when stored at -80°C. After reconstitution, prepare aliquots and store at -80°C. |
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Handling: | Avoid freeze/thaw cycles. |
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Emission maximum: | Excitation 485nm / Emission 520nm |
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Regulatory Status: | RUO - Research Use Only |
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HPLC analysis of BML-SE761.
DUB Assay: 50nM USP2 (Prod. No. BML-UW9850); 500nM Ubiquitin-Rhodamine; 50mM HEPES pH7.8, 0.5mM EDTA, 1mM DTT, and ovalbumin 1mg/ml.
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General Literature References
A sensitive fluorescence intensity assay for deubiquitinating proteases using ubiquitin-rhodamine110-glycine as substrate.: U. Hassiepen; Anal. Biochem.
371, 201 (2007),
Abstract;