Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Cell culture extracts, Tissue Extracts
Product nameCellular Calcineurin Phosphatase Activity Assay Kit (Colorimetric)
See all Calcineurin A kits
Sample typeCell culture extracts, Tissue Extracts
Assay typeEnzyme activity
Calcineurin Activity Assay Kit (Colorimetric) ab139464 is designed for measuring cellular calcineurin (CaN, PP2B) phosphatase activity. It employs a convenient 96-well microtiter-plate format with all reagents necessary for measuring calcineurin (PP2B) phosphatase activity in tissue/cellular extracts.
The detection of free-phosphate released in the calcineurin assay protocol is based on the classic Malachite green assay and offers the following advantages: non-radioactive; convenient 1-step detection; excellent sensitivity.
This kit includes human recombinant calcineurin as a positive control. The RII phosphopeptide substrate (Calcineurin Substrate), supplied with this calcineurin assay, is the most efficient and outstanding peptide substrate known for calcineurin.
Previously called Cellular Calcineurin Phosphatase Activity Assay Kit (Colorimetric).
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests 2X EGTA Buffer 1 x 1ml 96-well Clear Microplate (1/2 Volume) 2 units Active Calcineurin Enzyme 1 x 10µl Calcineurin Assay Buffer 1 x 20ml Calcineurin Substrate 1 x 1.5mg Calmodulin 1 x 100µl Desalting Column 1 unit Desalting Resin 1 x 1g Green Assay Reagent 1 x 20ml Lysis Buffer 1 x 40ml Okadaic Acid 1 x 325µl Phosphate Standard 1 x 0.5ml Protease Inhibitor Cocktail 1 x 2 tablets
FunctionCalcium-dependent, calmodulin-stimulated protein phosphatase. This subunit may have a role in the calmodulin activation of calcineurin. Dephosphorylates DNM1L, HSPB1 and SSH1.
Sequence similaritiesBelongs to the PPP phosphatase family. PP-2B subfamily.
Cellular localizationNucleus. Colocalizes with ACTN1 and MYOZ2 at the Z line in heart and skeletal muscle.
- Information by UniProt
- Alpha isoform formerly PPP2B
- Calcineurin A alpha
- Calcineurin A1
Example of standard curve obtained with ab139464.
Phosphatase activity from a freshly prepared Mouse brain extract. Prior to gel filtration the extract was diluted 1/1 in lysis buffer containing protease inhibitors. After gel filtration and prior to the phosphatase assay, the extract was diluted 1/25 in lysis buffer. The reaction was incubated for 30 min at 30°C.
Purified calcineurin positive control. The Calcineurin was incubated 1 hr at 30°C under various buffer conditions. The results demonstrate that Calcineurin activity is inhibited by the EGTA buffer, but not inhibited by 100 nM or 500 nM concentrations of okadaic acid.
ab139464 has been referenced in 3 publications.
- Zhang X et al. NULP1 Alleviates Cardiac Hypertrophy by Suppressing NFAT3 Transcriptional Activity. J Am Heart Assoc 9:e016419 (2020). PubMed: 32805187
- Zhu ZD et al. SOCE induced calcium overload regulates autophagy in acute pancreatitis via calcineurin activation. Cell Death Dis 9:50 (2018). PubMed: 29352220
- Triñanes J et al. Deciphering Tacrolimus-Induced Toxicity in Pancreatic ß Cells. Am J Transplant 17:2829-2840 (2017). PubMed: 28432716