Biological Information

Background Information:

Tissue plasminogen activator (tPA) is a trypsin-like serine protease produced and secreted primarily by vascular endothelial cells. Activation of tPA at the surface of blot clots causes the subsequent generation of plasmin that results in fibrin degradation. Aside from clot dissolution of thromboembolic events in coronary artery disease, acute myocardial infarction, ischemic stroke, peripheral arterial occlusive disease, venous thromboembolism and sepsis, tPA has been implicated in neurodegenerative diseases.

Family:

Protease

Sub Family:

Chymotrypsin

Class:

Serine Peptidase

Protein Name:

tPA

Uniprot Number:

P00750

Gene Name:

PLAT

Gene ID:

5327

Accession Number:

NM_000930

Organism:

Human

Construct Details:

Endogenous

Assay Information

Assay Type:

Biochemical

Assay Sub Type:

Enzymatic

Functional Mode:

Antagonist

Detection Method:

Spectrofluorimetry

Measured Response:

Fluorescence

Testing Information

Procedure Summary:

Native tPA from human melanoma cell is used. Test compound and/or vehicle is preincubated with the 160 U/mL enzyme in modified Tris-HCl buffer pH 7.6 for 15 minutes at 37˚C. The reaction is initiated by addition of 100 μM D-Ile-Pro-Arg-AMC for another 90-minute incubation period. Determination of the amount of AMC formed is read spectrofluorimetrically at Excitation: 360 nm, Emission: 465 nm. Compounds are screened at 10 μM.

Substrate:

100 μM D-Ile-Pro-Arg-AMC

Incubation:

90 min at 37°C

Control Inhibitor:

D-Phe-Pro-Arg-Chloromethylketone

Criteria For Significance:

≥ 50% of max stimulation or inhibition

Test Concentration / Dose:

Suggested concentration for initial testing is 10 μM

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

5311176

Name

PPACK

Measurement

0.063μM - IC50

Clinical Relevance

Therapeutic Area:

Cardiovascular

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

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