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Calcineurin

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Calcineurin is a calcium-dependent serine/threonine protein phosphatase. It plays a critical role in the regulation of cellular processes by dephosphorylating key target proteins.

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Lab products found in correlation

Calmodulin, by Enzo Life Sciences (2 mentions) Anti p ser 14 3 3 binding motif10, by Cell Signaling Technology (2 mentions) Anti p ser142 tfeb9, by Cell Signaling Technology (2 mentions)

2 protocols using calcineurin

1

Dephosphorylation of Activated TFEB

Cited 2 times
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Phosphorylated FLAG-TFEB was obtained by immunoprecipitation from fed HeLa cells. Protein-G beads containing FLAG-TFEB were incubated for 40 min at 30°C in 24 µl of corresponding phosphatase reaction buffer (20 mM HEPES, pH 7.5, 10 mM MgCl2, 2 mM DTT, protease inhibitors with 1 mM CaCl2 or 10 mM EGTA) containing purified Calcineurin (100U; ENZO Life Sciences) and Calmodulin (4 mM; ENZO Life Sciences). Samples were eluted by adding 8 µl of 4X Laemmli buffer, resolved by SDS–PAGE and analysed by immunoblotting. Antibodies anti-p-Ser 14-3-3 binding motif10 (link), anti-P-Ser142-TFEB9 (link) and TFEB (Cell Signaling Technologies) were used to detect phosphorylated and total TFEB forms, respectively.
Medina D.L., Di Paola S., Peluso I., Armani A., De Stefani D., Venditti R., Montefusco S., Scotto-Rosato A., Prezioso C., Forrester A., Settembre C., Wang W., Gao Q., Xu H., Sandri M., Rizzuto R., De Matteis M.A, & Ballabio A. (2015). Lysosomal calcium signaling regulates autophagy via calcineurin and TFEB. Nature cell biology, 17(3), 288-299.
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2

Dephosphorylation of Activated TFEB

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Phosphorylated FLAG-TFEB was obtained by immunoprecipitation from fed HeLa cells. Protein-G beads containing FLAG-TFEB were incubated for 40 min at 30°C in 24 µl of corresponding phosphatase reaction buffer (20 mM HEPES, pH 7.5, 10 mM MgCl2, 2 mM DTT, protease inhibitors with 1 mM CaCl2 or 10 mM EGTA) containing purified Calcineurin (100U; ENZO Life Sciences) and Calmodulin (4 mM; ENZO Life Sciences). Samples were eluted by adding 8 µl of 4X Laemmli buffer, resolved by SDS–PAGE and analysed by immunoblotting. Antibodies anti-p-Ser 14-3-3 binding motif10 (link), anti-P-Ser142-TFEB9 (link) and TFEB (Cell Signaling Technologies) were used to detect phosphorylated and total TFEB forms, respectively.
Medina D.L., Di Paola S., Peluso I., Armani A., De Stefani D., Venditti R., Montefusco S., Scotto-Rosato A., Prezioso C., Forrester A., Settembre C., Wang W., Gao Q., Xu H., Sandri M., Rizzuto R., De Matteis M.A, & Ballabio A. (2015). Lysosomal calcium signaling regulates autophagy via calcineurin and TFEB. Nature cell biology, 17(3), 288-299.
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