Synthesis of custom monoubiquitylated peptides designed from your protein of interest
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Choose the ubiquitylation site. . .
We’ll deliver a monoubiquitylated peptide!
We can provide custom monubiquitylated peptides from the sequence of your choosing. lsopeptide-linked ubiquitylated peptide reagents are produced using peptide synthesis technologies. Stepwise solid-phase synthesis and fragment condensation / chemical ligation approaches are employed in the synthesis of the ubiquitylated peptides. By utilizing these complementary methodologies for peptide synthesis, a large variety of native and labelled ‘isopeptide bond-linked’ ubiquitylated peptides can be generated for a variety of applications including:
• DUB substrates for screening
• Antigens for generation of site-specific ubiquitin antibodies
• Structural studies of ubiquitin-binding domain recognition
In contrast to traditional DUB substrates, like ubiquitin-AMC and ubiquitin-rhodamine, which measure cleavage of amide bonds, utilization of a monoubiquitylated peptide as a DUB substrate measures cleavage of an isopeptide bond and allows determination of the influence of sequence proximal to the ubiquitylated lysine on specificity. Profiling activity of a panel of DUBs against a monoubiquitylated histone H2B peptide and a monoubiquitylated PTEN peptide reveals interesting differences in specificity:
Assessment of DUB activity using monoubiquitylated peptides as a substrates. 2µM monoubiquitylated histone H2B peptide (red), 2µM monoubiquitylated PTEN peptide (blue), and 2µM ubiquitin-rhodamine were incubated with 50nM of each indicated DUB for 30 minutes at 37ºC. Isopeptidase cleavage of ubiquitin from the monoubiquitylated peptides was assessed by LC-MS and is represented as the percent substrate cleaved. Cleavage of ubiquitin-rhodamine was measured as the increase in rhodamine fluorescence at 535nm and is represented as the rate (RFU/sec) relative to USP2c. Monoubiquitylated PTEN and histone H2B peptides are available from stock.
Length: peptides of almost any length can be synthesized Quantity: µg to gram quantities Purity: choose purity levels from 80-98% Fluorescent modifications: examples include fluorescein, TAMRA, AlexaFluor®, or FRET pairs Affinity tags: examples include biotin, His6, FLAG®, V5 Ubiquitin options: peptides can be synthesized with wild-type or mutant ubiquitins, including lysine to arginine substitutions or unnatural amino acids
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