Biological Information
Background Information:
CD45 is a major cell surface glycoprotein expressed on nucleated hemapoietic cells. It is a member of the receptor-like transmembrane protein tyrosine phosphatase (PTPase) family. Studies have shown that CD45 is essential for T-cell receptor coupling couple to second messenger pathways involved in IL-2 production, and the antigen–mediated T-cell proliferation. One model of CD45 action postulates a role in dephosphorylation of lck kinase at the regulatory tyrosine, sensitizing the kinase response to T-cell receptor occupancy. Inhibitors of tyrosine phosphatase activity, therefore, may act as immunomodulators.
Family:
Phosphatase
Sub Family:
RTP
Class:
Protein Tyrosine
Protein Name:
CD45
Uniprot Number:
P08575
Gene Name:
PTPRC
Gene ID:
5788
Gene Aliases:
LCA|T200|GP180
Accession Number:
NM_002838
Organism:
Human
Construct Details:
Fragment (598-end), 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 CD45 expressed in E. coli is used. Test compound and/or vehicle is preincubated with 2.1 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 60 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:
60 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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