Biological Information

Background Information:

Calcineurin is a Calcineurin is a Ca2+/calmodulin-dependent protein phosphatase found in many cell types. It has been recently shown to be a key signaling enzyme in T-lymphocyte activation. It is also a major intracellular target for immunosuppressants such as the cyclosporin A/cyclophyllin complex. Inhibition of calcineurin, PP2B may be immunosuppressant, improve allograft survival in solid organ transplantation as well as useful in the treatment of some inflammatory diseases.

Family:

Phosphatase

Class:

Protein Ser/Thr

Protein Name:

PP2B

Uniprot Number:

Q08209

Protein Aliases:

calcineurin A alpha|protein phosphatase 2B, catalytic subunit, alpha isoform

Gene Name:

PPP3CA

Gene ID:

5530

Gene Aliases:

CNA1|PPP2B

Accession Number:

NM_000944 + NM_000945

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 PP2B expressed in E. coli is used. Test compound and/or vehicle is preincubated with 50 U/ml enzyme‡ for 15 minutes at 37°C in modified Tris-HCl buffer pH 7.2. The reaction is initiated by addition of 10 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:

10 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

PubChem ID

5566

Name

Trifluoperazine

Measurement

100μM - IC50

PubChem ID

61671

Name

Trisodium vanadate

Measurement

300μM - IC50

PubChem ID

16760572

Name

MpV(Pic)

Measurement

3.6μM - IC50

PubChem ID

329770925

Name

(NH4)6Mo7O24

Measurement

25μM - IC50

PubChem ID

24277738

Name

Suramine

Measurement

5.3μM - IC50

Clinical Relevance

Adverse / Beneficial Action:

Inhibition of calcineurin, PP2B, may be immunosuppressant, improve allograft survival in solid organ transplantation, as well as useful in the treatment of some inflammatory diseases

Additional Information

Brand:

LeadHunter

Testing Location:

Taiwan - Taipei

RELATED SOLUTIONS

Item: PP89