ActivX Desthiobiotin-GTP Probe

Description

Thermo Scientific Pierce ActivX Desthiobiotin-GTP Probe covalently label the active site of GTPases and GTPase subunits of G-protein coupled receptors enabling their selective enrichment thereby allowing identification and profiling of target enzyme classes across samples or to assess the specificity and affinity of enzyme inhibitors.

Features of ActivX Desthiobiotin-GTP Probe:

• Specific—label only the conserved active-site lysines of nucleotide-binding proteins
• Compatible—use for in vitro labeling of GTPase enzymes derived from cells or tissues.
• Flexible—use with Western blot or mass spectrometry (MS) workflows

Applications of ActivX Desthiobiotin-GTP Probe:
• Broad enrichment of GTP-binding proteins from tissues, cells and sub-cellular proteomes
• Enrichment of enzymes based on function
• Profiling of dozens to hundreds of inhibitor targets

ActivX Desthiobiotin-GTP Probe structure consists of a modified biotin attached to the nucleotide by a labile acyl-phosphate bond. After removal of GTP or GDP nucleotides from enzymes, the desthiobiotin-GTP probe can be used to covalently modify conserved lysine residues in the GTPase nucleotide-binding site. Desthiobiotin-GTP can selectively enrich, identify and profile target enzyme classes in samples. Pre-incubation of samples with small-molecule inhibitors that compete with active-site probes can be used to determine inhibitor binding affinity and target specificity.

Assessment of active-site labeling can be accomplished by either Western blot or mass spectrometry (MS). For the Western blot workflow, desthiobiotin-labeled proteins are enriched for SDS-PAGE analysis and subsequent detection with specific antibodies. For the MS workflow, desthiobiotin-labeled proteins are reduced, alkylated and enzymatically digested to peptides. Only the desthiobiotin-labeled, active-site peptides are enriched for analysis by LC-MS/MS. Both workflows can be used for determining inhibitor target binding, but only the MS workflow can identify global inhibitor targets and off-targets.

More Product Data
• GTPase enrichment using a new active-site probe

Related Products
Pierce™ GTPase Enrichment Kit with GTP Probe

Figures

Specifications

Assay:	Protein Quantitation Assay
Detection Method:	Chemiluminescent, Mass Spectrometry, Western Blot
Label Type:	Biotin & Other Haptens
Product Line:	ActivX™
Substrate Properties:	Chemical Substrate
Substrate Type:	GTPase Substrate
Target Enzyme:	Kinases (Multiple), Phosphodiesterases, ATPases, GTPases, Phosphatases
Technique:	Pull-Down
Format:	Tube(s), Spin Column
Quantity:	16 x 12.9µg

Contents & storage