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

PubChem ID

61671

Name

Trisodium vanadate

Measurement

0.016μM - IC50

PubChem ID

91826558

Name

Na2MoO4.7H2O

Measurement

0.27μM - IC50

PubChem ID

329770925

Name

(NH4)6Mo7O24

Measurement

0.036μM - IC50

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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