Biological Information
Background Information:
PTP1C (also known as HCP, SHP and SH-PTP1) is a non-transmembrane type protein tyrosine phosphatase that contains two src homology-2 (SH2) domains. The SH2 domain is believed to have dual roles: as a facilitator promoting interactions between cytoplasmic signalling molecules and specific phosphotyrosyl residues on activated (i.e., autophosphorylated) growth factor receptors, and as a self regulator of phosphatase activity. PTP1C is expressed in cells of all hematopoetic lineages. Its function has recently been linked to immune regulatory activity. This assay may be used to identify inhibitors and potential therapeutic drugs for autoimmune diseases.
Family:
Phosphatase
Sub Family:
Non-Receptor
Class:
Protein Tyrosine
Protein Name:
SHP1
Uniprot Number:
P29350
Gene Name:
PTPN6
Gene ID:
5777
Gene Aliases:
HCP|HCPH|PTP-1C|SHP-1|SHP1
Accession Number:
NM_002831
Organism:
Human
Construct Details:
Fragment (243-541), Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human recombinant protein phosphatase SHP-1 expressed in E. coli is used. Test compound and/or vehicle is preincubated with 2 U/ml enzyme‡ for 15 minutes at 37°C in modified HEPES buffer pH 7.2. The reaction is initiated by addition of 100 μM DiFMUP for 30 minutes. Determination of the amount of DiFMU formed is read spectrofluorimetrically at 358 nm/450 nm. Compounds are screened at 10 μM.
Substrate:
100 µM DiFMUP
Incubation:
30 min at 37°C
Control Inhibitor:
(NH₄)₆Mo₇O₂₄
Criteria For Significance:
≥ 50% of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Test Sample Requirements:
Minimum for 1) Screen: 25 μl of 10 mM stock -OR- 1 mg (pre-weighed). 2) Dose Response: 60 μl of 10 mM stock -OR- 1 mg (pre-weighed).
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei
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