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Ab154650

Manufactured by Abcam
Sourced in United States

Ab154650 is a lab equipment product offered by Abcam. It is a device used for scientific research purposes. The core function of this product is to perform a specific task in a laboratory setting.

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2 protocols using ab154650

1

Evaluation of SELENOK in Melanoma Cells

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Lysates from primary melanocytes were purchased from Sciencell Research Laboratories (Carlsbad, CA) and NCI-60 validated human melanoma cell lines obtained from the University of Hawaii Cancer Center included SK-Mel2, SK-Mel28, and MALME-3M. These cell lines were cultured in RPMI media with 10% fetal bovine serum and 1% Antibiotic-Antimycotic (all from GIBCO/Thermo Fisher). Primary antibodies for western blots included rabbit monoclonal anti-SELENOK (Epigemonics, Inc., custom antibody), anti-IP3R1 (Santa Cruz Biotechnology, sc-271197), anti-GAPDH (Santa Cruz Biotechnology, sc-47724), anti-Prom1 (Cell Signaling, 58605). Antibodies from Abcam included anti-TRP2 (ab74073), anti-calcineurin A and B (ab137335, ab154650), and anti-calmodulin (ab 105498, respectively). Western blot secondary antibodies were purchased from Li-Cor Technologies and immunofluorescence secondary Alexafluor595 antibody from Thermo. The transgene encoding full-length SELENOK in pcDNA3.1+ (Invitrogen) has been previously described [11 (link)].
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2

Colorimetric Assay for Calcineurin

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Lyophilized CNAΔ316, CNB standard, CNB test sample (Beijing Normal University), p-nitrophenylphosphate (pNPP) (34047, Thermo Fisher Science, USA), bovine serum albumin (BSA) (ST023 Beyotime, Shanghai), DLdithiothreitol (DTT) (ST040, Beyotime, Shanghai), and anti-CNB polyclonal antibody (ab154650, Abcam, USA) were used.
Testing medium comprised 50 mmol L 1 Tris-HCl (pH 7.4), 0.2 mg mL 1 BSA, 20 mmol L 1 pNPP, 1 mmol L 1 DTT, 1 mmol L 1 MnCl 2 , and 1 mmol L 1 CaCl 2 . Microtiter plate reader (Spectrum plus 5, Molecular Device, USA) was used in the assays.
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