Technology Portfolio

DCP-Bio1: Cell-compatible Probe for Analyzing Protein Oxidation

DCP-Bio1 is a dimedone-based, sulfenic acid-reactive compound containing a biotin tag, making it compatible with several techniques and forms of analysis for identifying redox-regulated proteins and their specific sites of modification.

Tool Overview

Ref #: WFU 13-17
  • Contains biotin tag as affinity handle; no need for ‘click’ chemistry before detection or protein isolation
  • Cell-permeable
  • Stable, reproducible covalent binding to cysteine sulfenic acid (-SOH) at pH 6.0 ­– pH 8.0
  • Ester linkage stays intact under physiological conditions, but allows for efficient hydrolysis using NH4OH
    (Doesn’t hydrolyze under physiological conditions)
  • Effective in in vitro and in vivostudies
  • Compatible with Western Blotting, ELISA, and Affinity Isolation and cell-imaging techniques

Redox-sensitive cysteine residues in proteins may serve as important components of oxidative signaling or sensors of oxidative stress. Cysteine sulfenic acid modification is an emerging area of interest for those studying biological signal transduction within the cell.

Cysteine sulfenic acid formation in proteins results from the oxidative modification of susceptible cysteine residues by signaling-relevant cellular oxidants such as hydrogen peroxide, lipid hydroperoxides and peroxynitrite. These sulfenic acid-modified proteins can be identified by their ability to form adducts with dimedone, but this reagent provides no spectral or affinity tag.

DCP-Bio1 can be used to detect the formation of cysteine sulfenic acid as a common initial step in the redox regulation of proteins, and with the presence of a biotin label, DCP-Bio1 is compatible with several techniques and forms of analysis post-labeling.

Added to intact cells, lysates or protein samples to trap oxidized cysteines chemically, this reagent can be used to study single proteins in isolation or in the context of cells or proteomics-wide protein oxidation in systems of interest.

Full Name: DCP-Bio1; 3-(2,4-dioxocyclohexyl)propyl 5-((3aR,6S,6aS)-hexahydro-2-oxo-1H-thieno[3,4-d]imidazol-6-yl)pentanoate

Chemical Formula: C19H28N2O5S

Molecular Weight: 396.5 g/mol

Format: Solid

Purity: >98% pure

Solubility: At least 500 mM in DMSO, at least 5 mg/ml in acetonitrile

Stability: Stable > 6 months at -20° C

Source: Synthetic

Spectral Information: No visible absorbance

Storage: Room temperature for short term, -20° C for long term

Comments: Stock solution in DMSO can be added to buffers. Can dissolve in acetonitrile to aliquot and redry.

  • Leslie B. Poole, PhD, Department of Biochemistry
  • Bruce King, PhD, Department of Chemistry
  • Cristina M. Furdui, PhD, Department of Internal Medicine