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Safe and non-radioactive assay
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Convenient 1-step detection
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Excellent sensitivity
A complete colorimetric assay kit for measuring cellular calcineurin (PP2B) phosphatase activity. It employs a convenient 96-well plate format with all reagents necessary for measuring calcineurin (PP2B) phosphatase activity in tissue/cellular extracts PLUS human recombinant calcineurin is included as a positive control. The RII phosphopeptide substrate, supplied with this kit is the most efficient and selective peptide known for calcineurin. The detection of free-phosphate released is based on the classic malachite green assay.
CaN Positive Control. Purified calcineurin (BML-SE163-9090) positive control. The CaN was incubated 1 hr at 30°C under various buffer conditions. The results demonstrate that CaN activity is inhibited by the EGTA buffer, but not inhibited by 100nM or 500nM concentrations of okadaic acid
Cellular CaN Assay Using Mouse Brain 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. Well contents were as in manual. The reaction was incubated for 30 min at 30°C.
BIOMOL GREEN™ Phosphate Standard Curve
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Product Details
| Applications: | Colorimetric detection
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| Use/Stability: | Store all components except the microtiter plate at 80°C for the highest stability. The calcineurin enzyme component SE163-9090 must be handled particularly carefully in order to retain maximal enzymatic activity. Thaw it quickly in a RT water bath or by rubbing between fingers, then immediately store on an ice bath. The remaining unused enzyme should be quickly refrozen by placing at -80°C. To minimize the number of freeze/thaw cycles, aliquot the calcineurin into separate tubes and store at -80°C. |
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| Shipping: | Dry Ice |
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| Long Term Storage: | -80°C |
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| Contents: |
Calcineurin enzyme, Calmodulin (human, recombinant), Substrate (RII phosphopeptide), 2x assay buffer, EGTA Buffer, Lysis Buffer, Protease inhibitor cocktail, Biomol Green™ Reagent, Phosphate standard, Okadaic acid, Desalting column and resin, ½-Volume microplate |
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| Regulatory Status: | RUO - Research Use Only |
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| Compatibility: | This product is compatible with the Absorbance 96 Plate Reader.
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Product Literature References
A novel implant surface modification mode of Fe3O4-containing TiO2 nanorods with sinusoidal electromagnetic field for osteoblastogenesis and angiogenesis: R. Ren, et al.; Mater. Today Bio
19, 100590 (2023),
Abstract;
Heme oxygenase-1 gene delivery for altering high mobility group box-1 protein in pancreatic islet: M.J. Kim, et al.; J. Control Release
343, 326 (2022),
Abstract;
Involvement of mechano-sensitive Piezo1 channel in the differentiation of brown adipocytes: M. Kenmochi, et al.; J. Physiol. Sci.
72, 13 (2022),
Abstract;
JCI‑20679 suppresses the proliferation of glioblastoma stem cells by activating AMPK and decreasing NFATc2 expression levels: S. Ando, et al.; Mol. Med. Rep.
26, 238 (2022),
Abstract;
The optimized core peptide derived from CABIN1 efficiently inhibits calcineurin-mediated T-cell activation: S. Lee, et al.; Exp. Mol. Med.
54, 613 (2022),
Abstract;
A yeast-based screening system identified bakkenolide B contained in Petasites japonicus as an inhibitor of interleukin-2 production in a human T cell line: S. Uesugi, et al.; Biosci. Biotechnol. Biochem.
85, 2153 (2021),
Abstract;
Calcineurin Activation by Prion Protein Induces Neurotoxicity via Mitochondrial Reactive Oxygen Species: J.H. Moon, et al.; Oxid. Med. Cell. Longev.
2021, 5572129 (2021),
Abstract;
Calcineurin activity is increased in Charcot-Marie-Tooth 1B demyelinating neuropathy: M. Sidoli, et al.; J. Neurosci.
1523, 2384 (2021),
Abstract;
Calcineurin, Calcium-Dependent Serine-Threonine Phosphatase Activation by Prion Peptide 106-126 Enhances Nuclear Factor-κB-Linked Proinflammatory Response through Autophagy Pathway: J.M. Hong, et al.; ACS Chem. Neurosci.
12, 3277 (2021),
Abstract;
Decreased IFT88 expression with primary cilia shortening causes mitochondrial dysfunction in cisplatin-induced tubular injury: R. Fujii, et al.; Am. J. Physiol. Renal Physiol.
321, F278 (2021),
Abstract;
Acylated Ghrelin Protects the Hearts of Rats from Doxorubicin-Induced Fas/FasL Apoptosis by Stimulating SERCA2a Mediated by Activation of PKA and Akt: A.A. Shati, et al.; Cardiovasc. Toxicol.
19, 529 (2019),
Abstract;
Fas/FasL-mediated cell death in rat's diabetic hearts involves activation of calcineurin/NFAT4 and is potentiated by a high-fat diet rich in corn oil: R.A. Eid, et al.; J. Nutr. Biochem.
68, 79 (2019),
Abstract;
Redox signals at the ER–mitochondria interface control melanoma progression: X. Zhang, et al.; EMBO J.
38, e100871 (2019),
Abstract;
Full Text
AKAP150 involved in Paclitaxel-Induced Neuropathic Pain via Inhibiting CN/NFAT2 pathway and downregulating IL-4: B. Nie, et al.; Brain Behav. Immun.
68, 158 (2018),
Abstract;
Angiotensin II Type I Receptor Agonistic Autoantibody Induces Podocyte Injury via Activation of the TRPC6- Calcium/Calcineurin Pathway in Pre-Eclampsia: Y. Yu, et al.; Kidney Blood Press Res.
43, 1666 (2018),
Abstract;
Human immunodeficiency virus Tat impairs mitochondrial fission in neurons: S.J. Rozzi, et al.; Cell Death Discov.
4, 8 (2018),
Abstract;
Full Text
Isx9 Regulates Calbindin D28K Expression in Pancreatic β Cells and Promotes β Cell Survival and Function: J.B. Pujol, et al.; Int. J. Mol. Sci.
19, 2542 (2018),
Abstract;
Full Text
RANK Deficiency Ameliorates Podocyte Injury by Suppressing Calcium/Calcineurin/ NFATc1 Signaling: S. Liang, et al.; Kidney Blood Press. Res.
43, 1149 (2018),
Abstract;
Selenoprotein K deficiency inhibits melanoma by reducing calcium flux required for tumor growth and metastasis: M.P. Marciel, et al.; Oncotarget
9, 13407 (2018),
Abstract;
Full Text
Cardiac-directed expression of a catalytically inactive adenylyl cyclase 6 protects the heart from sustained β-adrenergic stimulation: M.H. Gao, et al.; PLoS One
12, e0181282 (2017),
Application(s): Activity assay using left ventricle chamber of mouse,
Abstract;
Full Text
Increased PKC activity and altered GSK3β/NMDAR function drive behavior cycling in HINT1-deficient mice: bipolarity or opposing forces: J. Garzon-Nino, et al.; Sci. Rep.
7, 43468 (2017),
Abstract;
Full Text
IP3R-mediated Ca2+ signals govern hematopoietic and cardiac divergence of Flk1+ cells via the calcineurin-NFATc3-Etv2 pathway: Y.J. Wang, et al.; J. Mol. Cell. Biol.
9, 274 (2017),
Abstract;
Mitochondrial Abnormality Facilitates Cyst Formation in Autosomal Dominant Polycystic Kidney Disease: Y. Ishimoto, et al.; Mol. Cell. Biol.
37, e00337-17 (2017),
Abstract;
Full Text
Obesity-associated cardiac pathogenesis in broiler breeder hens: Pathological adaption of cardiac hypertrophy: C.Y. Chen, et al.; Poult. Sci.
96, 2428 (2017),
Abstract;
Pulsed electromagnetic field inhibits RANKL-dependent osteoclastic differentiation in RAW264.7 cells through the Ca2+-calcineurin-NFATc1 signaling pathway: J. Zhang, et al.; Biochem. Biophys. Res. Commun.
482, 289 (2017),
Abstract;
Reduced cortical excitatory synapse number in APOE4 mice is associated with increased calcineurin activity: A.L. Neustadtl, et al.; Neuroreport
28, 618 (2017),
Abstract;
Full Text
Subcellular Distribution of HDAC1 in Neurotoxic Conditions Is Dependent on Serine Phosphorylation: Y. Zhu, et al.; J. Neurosci.
37, 7547 (2017),
Abstract;
Full Text
TRAF6 Mediates Suppression of Osteoclastogenesis and Prevention of Ovariectomy-Induced Bone Loss by a Novel Prenylflavonoid: E.M. Tan, et al.; J. Bone Miner. Res.
32, 846 (2017),
Abstract;
CRTC1 nuclear localization in the hippocampus of the pilocarpine-induced status epilepticus model of temporal lobe epilepsy: D. Dubey & B.E. Porter; Neuroscience
320, 57 (2016),
Application(s): Measure calcineurin levels in hippocampal lysates,
Abstract;
KRAS mutation leads to decreased expression of regulator of calcineurin 2, resulting in tumor proliferation in colorectal cancer: H. Niitsu, et al.; Oncogenesis
5, e253 (2016),
Application(s): Calcineurin activity assay, cell lysates,
Abstract;
Full Text
New strategy of tacrolimus administration in animal model based on tacrolimus-loaded microspheres: J.J. Zamorano-Leon, et al.; Transplant Immunol.
36, 9 (2016),
Abstract;
Differential effects of short-term β agonist and growth hormone treatments on expression of myosin heavy chain IIB and associated metabolic genes in sheep muscle: K.M. Hemmings, et al.; Animal
9, 285 (2015),
Application(s): Use with skeletal muscle homogenates,
Abstract;
Full Text
Myocyte enhancer factor (MEF)-2 plays essential roles in T-cell transformation associated with HTLV-1 infection by stabilizing complex between Tax and CREB: P. Jain, et al.; Retrovirology
12, 23 (2015),
Application(s): Assay,
Abstract;
Full Text
Regulation of GAP43/calmodulin complex formation via calcineurin-dependent mechanism in differentiated PC12 cells with altered PMCA isoforms composition: T. Boczek, et al.; Mol. Cell. Biochem.
407, 251 (2015),
Application(s): Assay using cell lysate,
Abstract;
Full Text
The Ca2+-activated cation channel TRPM4 is a negative regulator of angiotensin II-induced cardiac hypertrophy: M. Kecskés, et al.; Basic Res. Cardiol.
110, 43 (2015),
Application(s): Assay using mouse heart supernatant,
Abstract;
Full Text
TRPV6 channel modulates proliferation of insulin secreting INS-1E beta cell line: M. Skrzypski, et al.; Biochim. Biophys. Acta
1853, 3202 (2015),
Application(s): Calcineurin activity in rat pancreatic cells,
Abstract;
Calcineurin suppresses AMPK-dependent cytoprotective autophagy in cardiomyocytes under oxidative stress: H. He, et al.; Cell Death Dis.
5, e997 (2014),
Application(s): Activity measured in rat cardiomyocyte cellular extracts,
Abstract;
Full Text
Calcineurin upregulates local Ca2+ signaling through ryanodine receptor-1 in airway smooth muscle cells: C. Savoia et al.; Am. J. Physiol. Lung Cell. Mol. Physiol.
307, 781 (2014),
Abstract;
Full Text
Calcineurin activity is required for cardiac remodelling in pregnancy: E. Chung, et al.; Cardiovasc. Res.
100, 402 (2013),
Abstract;
Full Text
Calcium phosphatase calcineurin influences tau metabolism: C.M. Karch, et al.; Neurobiol. Aging
34, 374 (2013),
Application(s): Calcineurin phosphatase activity in extracted brain and cell lysates,
Abstract;
IF1 limits the apoptotic-signalling cascade by preventing mitochondrial remodelling: D. Faccenda, et al.; Cell Death Differ.
20, 686 (2013),
Application(s): Calcineurin phosphatase activity in HeLa cells and mouse embryonic fibroblasts,
Abstract;
Intracellular two-phase Ca2+ release and apoptosis controlled by TRP-ML1 channel activity in coronary arterial myocytes: M. Xu, et al.; Am. J. Physiol. Cell Physiol.
304, C458 (2013),
Abstract;
Full Text
The R740S mutation in the V-ATPase a3 subunit increases lysosomal pH, impairs NFATc1 translocation, and decreases in vitro osteoclastogenesis: I. Voronov, et al.; J. Bone Miner. Res.
28, 108 (2013),
Abstract;
Leptin-induced cardiomyocyte hypertrophy reveals both calcium-dependent and calcium-independent/RhoA-dependent calcineurin activation and NFAT nuclear translocation: V. Rajapurohitam, et al.; Cell Signal.
24, 2283 (2012),
Application(s): Calcineurin phosphatase activity in rat cardiac myocytes,
Abstract;
Molecular mechanisms underlying effects of neural stem cells against traumatic axonal injury: E. Wang, et al.; J. Neurotrauma
29, 295 (2012),
Application(s): Calcineurin phosphatase activity in neural stem cells,
Abstract;
Sustained postexercise increases in AS160 Thr642 and Ser588 phosphorylation in skeletal muscle without sustained increases in kinase phosphorylation: G.G. Schweitzer, et al.; J. Appl. Physiol. (1985)
113, 1852 (2012),
Abstract;
Full Text
The antimicrobial peptide, LL-37, inhibits in vitro osteoclastogenesis: C. Supanchart, et al.; J. Dent. Res.
91, 1071 (2012),
Application(s): Calcineurin phosphatase activity in human osteoclast progenitor cells,
Abstract;
Beneficial effects of adenylyl cyclase type 6 (AC6) expression persist using a catalytically inactive AC6 mutant: M.H. Gao, et al.; Mol. Pharmacol.
79, 381 (2011),
Application(s): Calcineurin phosphatase activity in adult rat cardiac myocytes,
Abstract;
Calcineurin/nuclear factor of activated T cells (NFAT) signaling in cobalt-chromium-molybdenum (CoCrMo) particles-induced tumor necrosis factor-α (TNFα) secretion in MLO-Y4 osteocytes: V. Orhue, et al.; J. Orthop. Res.
29, 1867 (2011),
Application(s): Calcineurin phosphatase activity in osteocytes,
Abstract;
Ginseng inhibits cardiomyocyte hypertrophy and heart failure via NHE-1 inhibition and attenuation of calcineurin activation: J. Guo, et al.; Circ. Heart Fail.
4, 79 (2011),
Application(s): Calcineurin phosphatase activity in rat cardiac myocytes,
Abstract;
Regulator of calcineurin 1 (Rcan1) is required for the development of pulmonary eosinophilia in allergic inflammation in mice: Y.J. Yang, et al.; Am. J. Pathol.
179, 1199 (2011),
Application(s): Calcineurin phosphatase activity in murine bone marrow cells,
Abstract;
A conserved calcineurin-binding motif in human T lymphotropic virus type 1 p12I functions to modulate nuclear factor of activated T cell activation: S.J. Kim, et al.; J. Biol. Chem.
278, 15550 (2003),
Application(s): Jurkat T cells,
Abstract;
Full Text
Targeted inhibition of calcineurin prevents agonist-induced cardiomyocyte hypertrophy: T. Taigen, et al.; PNAS
97, 1196 (2000),
Application(s): Rat cardiomyocytes,
Abstract;
Full Text
General Literature References
Overexpression and purification of human calcineurin alpha from Escherichia coli and assessment of catalytic functions of residues surrounding the binuclear metal center: A. Mondragon et al.; Biochemistry
36, 4934 (1997),
Abstract;
Characterization and kinetic analysis of the intracellular domain of human protein tyrosine phosphatase beta (HPTP beta) using synthetic phosphopeptides: K. W. Harder et al.; Biochem. J.
298 , 395 (1994),
Abstract;
Dephosphorylation of phosphopeptides by calcineurin (protein phosphatase 2B): A. Donella-Deana et al.; Eur. J. Biochem.
219, 109 (1994),
Abstract;
Nonradioactive assay for protein phosphatase 2B (calcineurin) activity using a partial sequence of the subunit of cAMP-dependent protein kinase as substrate: A. Enz et al.; Anal. Biochem.
216, 147 (1994),
Abstract;
Use of fluorinated tyrosine phosphates to probe the substrate specificity of the low molecular weight phosphatase activity of calcineurin: B. Martin et al.; J. Biol. Chem.
260, 14932 (1985),
Abstract;
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