Alkyne β-ketoester (Alk-β-KE)

The Alkyne β-ketoester probe (Alk-β-KE) can be utilized as a robust chemical probe for the labeling and analysis of sulfenic acid (-SOH) modified proteins. This probe is ideal for in vitro and in vivo applications and mass spectrometric (MS) analyses. Alk-β-KE is non-dimedone based probe that is customizable through the addition of biotin or other tags. The tags can be subsequently cleaved using NH₂OH making this probe ideal for quantitive analysis via mass spectrometry (MS).

Features:

Stable, reproducible binding to cysteine sulfenic acid (-SOH) at physiological pH; increased reactivity under basic conditions
Highly customizable. Biotin or other tags can be added to Alk-β-KE through the robust “click” reaction. The tags can be then removed using NH₂OH to yield a derivative amenable for quantitative analysis (via MS).
Great for in vitro and in vivo applications
Compatible with MS, WB, ELISA, and Affinity Isolation

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.

From the laboratory of Cristina M. Furdui, PhD, Wake Forest School of Medicine.

Part of The Investigator's Annexe program.

Catalog Number	Product	Description	Price	Order
EE1001	Alk-β-KE, 1mg	1mg	Price: $214.00	Qty:
EE1002	Alk-β-KE, 5mg	5mg	Price: $640.00	Qty:
EE1003	Alk-β-KE, 10mg	10mg	Price: $1,125.00	Qty:

Compound Name:	Alkyne β-ketoester probe (Alk-β-KE)
Chemical Formula:	C₈H₁₀O₃
Molecular Weight:	154.1632
Physical Form:	yellow oil
Purity:	> 95% by NMR
Stability:	> 6 months at -20C
Storage Conditions:	Store at -20C
Shipped:	Cold pack

Comments

Reaction of –SOH in proteins with Alk-β-KE. Proteins labeled by Alk-β-KE can be conjugated with reporter tags via click reaction. The reporter tag can then be removed using NH₂OH. The protein structure shown is C165S AhpC and was generated using Swiss PDB Viewer 4.0.1 based on the PDB entry 3EMP.

Adapted from: Qian, J., Wani, R., Klomsiri, C., Poole, L.B., Tsang, A.W., and Furdui, C.M. A simple and effective strategy for labeling cysteine sulfenic acid in proteins by utilization of β-ketoesters as cleavable probes. Chem. Commun., 48, 4091–4093 (2012).

ESI-TOF MS spectra showing the labeling of C165S AhpC–SOH by Alk-β-KE (adduct peak: 20 752 amu) (A); biotinylation of AhpC–Alk-β-KE adduct via click reaction (adduct peak: 21 549 amu) (B); and removal of the biotin moiety after NH2OH treatment (adduct peak: 20 697 amu) (C).>/p>

Adapted from: Qian, J., Wani, R., Klomsiri, C., Poole, L.B., Tsang, A.W., and Furdui, C.M. A simple and effective strategy for labeling cysteine sulfenic acid in proteins by utilization of β-ketoesters as cleavable probes. Chem. Commun., 48, 4091–4093 (2012).

References

Qian, J., Wani, R., Klomsiri, C., Poole, L.B., Tsang, A.W., and Furdui, C.M. A simple and effective strategy for labeling cysteine sulfenic acid in proteins by utilization of β-ketoesters as cleavable probes. Chem. Commun., 48, 4091–4093 (2012).

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