Biological Information

Background Information:

Cell Division Cycle 25B (CDC25B) is a member of the CDC25 family of dual specificity protein tyrosine phosphatases that dephosphorylate and activate cyclin-dependent kinases (CDKs). Mammalian cells express at least three dual phosphatase Cdc25 homologues, named A, B, and C. CDC25B has been proposed to activate the CDK1/cyclin B1 complex to advance the cell cycle from G2 to M.

Family:

Phosphatase

Sub Family:

Dual Specificity

Class:

Protein Tyrosine

Protein Name:

CDC25B

Uniprot Number:

P30305

Gene Name:

CDC25B

Gene ID:

994

Accession Number:

NM_021873

Organism:

Human

Construct Details:

Full Length, 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 CDC25B expressed in E. coli is used. Test compound and/or vehicle is preincubated with 0.61 μg/ml enzyme‡ for 15 minutes at 37°C in modified Tris-HCl buffer pH 8.5. The reaction is initiated by addition of 15 μM 3-O-methylfluorescein phosphate for 60 minutes. Determination of the amount of 3-O-methylfluorescein formed is read spectrofluorimetrically at 475 nm/530 nm. Compounds are screened at 10 µM.

Substrate:

15 µM 3-O-methylfluorescein phosphate

Incubation:

60 min at 37°C

Control Inhibitor:

NSC 663284

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

0.12μM - IC50

PubChem ID

379077

Name

NSC 663284

Measurement

0.09μM - IC50

PubChem ID

16078950

Name

BN82002 hydrochloride

Measurement

6.8μM - IC50

PubChem ID

329770925

Name

(NH4)6Mo7O24

Measurement

10μM - IC50

Clinical Relevance

Therapeutic Area:

Oncology/Immuno-Oncology

Adverse / Beneficial Action:

CDC25 is an oncogene and its inhibition may be useful in reducing tumor cell proliferation.

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei