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Calcineurin cellular activity assay kit

BML-AK816

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Provides everything needed to assay calcineurin cellular activity, including recombinant calcineurin control

Safe and non-radioactive assay
Convenient 1-step detection
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.

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Calcineurin cellular activity assay kit Standard curve — BIOMOL GREEN™ Phosphate Standard Curve

Calcineurin cellular activity assay kit Fig3 — 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

Calcineurin cellular activity assay kit Fig2 — 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.

Product Details

Application	Colorimetric detection
Compatibility	This product is compatible with the Absorbance 96 Plate Reader.
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

Handling & Storage

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.
Long Term Storage	-80°C
Shipping	Dry Ice

Regulatory Status	RUO – Research Use Only

Soluble β-Amyloid Oligomers Selectively Upregulate TRPC3 in Excitatory Neurons via Calcineurin-Coupled NFAT: Wang, Z., Ding, D., et al.; Cells 14, 843 (2025), Abstract
TRPV4 Channels Promote Pathological, but Not Physiological, Cardiac Remodeling through the Activation of Calcineurin/NFAT and TRPC6: Yañez-Bisbe, L., Moya, M., et al.; Int. J. Mol. Sci. 25, (2024), Abstract
TRPC3 suppression ameliorates synaptic dysfunctions and memory deficits in Alzheimer’s disease: Wang, J., Chen, L., et al.; bioRxiv , (2024)
Evaluation of hepatic AMPK, mTORC1, and autophagy-lysosomal pathway in cows with mild or moderate fatty live: X. Du, et al.; J. Dairy Sci. 107, 3269 (2024), Abstract
Melatonin-mediated calcineurin inactivation attenuates amyloid beta-induced apoptosis: Hong, J. M., Munna, A. N., et al.; IBRO Neurosci. Rep. 16, 336 (2024), Abstract
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
Calcineurin suppresses rat H9c2 cardiomyocyteprotective autophagy under chronic intermittent hypoxia by downregulating the AMPK pathway: C. Ke, et al.; Exp. Cell Res. 433, 113850 (2023), Abstract
Melatonin inhibits Japanese encephalitis virus replication and neurotoxicity via calcineurin-autophagy pathways: Moon, J. H., Hong, J. M., et al.; BMC Neurosci. 24, 59 (2023), Abstract
PAAN/MIF nuclease inhibition prevents neurodegeneration in Parkinson’s disease: Park, H., Kam, T. I., et al.; Cell 185, 1943 (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