Cells were fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) (09154-85; Nacalai Tesque) for 3 min and then incubated with an alkaline phosphatase staining solution at 37°C for 10–15 min. The staining solution consisted of 0.6 mg/mL Fast Red TR Salt [hemi(zinc chloride) salt, F8764; Sigma-Aldrich, St. Louis, MO, United States], 0.1 mg/mL naphthol phosphate (23821-24; Nacalai Tesque), 0.7 mM N,N-dimethylformamide (13016-65; Nacalai Tesque), 0.2 mM MgCl2 (20908-65; Nacalai Tesque), and 0.1 mM Tris-HCl (pH 8.5). The staining reaction stopped after ∼15 min due to fixation with the 4% paraformaldehyde solution. More details are described in our previous reports (Donai et al., 2013 (link)).
Naphthol phosphate
Manufactured by Nacalai Tesque
Sourced in United States
Naphthol phosphate is a chemical compound used as a substrate in various laboratory assays and diagnostic tests. It is a colorless, crystalline solid that undergoes enzymatic hydrolysis to produce a colored product, allowing for the detection and quantification of specific analytes. Naphthol phosphate is widely used in biochemical and clinical applications, but its detailed description and intended use should be provided by subject matter experts to ensure accuracy and avoid potential misinterpretation.
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Lab products found in correlation
2 protocols using naphthol phosphate
1
Alkaline Phosphatase Staining Protocol
2
Alkaline Phosphatase Staining of Stem Cells
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The pv-iPSCs were fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) (09154-85; Nacalai Tesque) for 3 min. The alkaline phosphatase (ALP) enzymatic activity was detected based on the protocol described in our previous study (4) . In brief, the staining solution, consisting of 0.6 mg/ml Fast Red TR Salt [hemi(zinc chloride) salt, F8764; Sigma-Aldrich, St. Louis, MO, USA], 0.1 mg/ml naphthol phosphate (23821-24; Nacalai Tesque), 0.7 mM N,N-dimethylformamide (13016-65; Nacalai Tesque), 0.2 mM MgCl 2 (20908-65; Nacalai Tesque), and 0.1 mM Tris-HCl (pH 8.5), was mixed with the fixed cells, and incubated at 37°C for 10 min. After the staining reaction had run its course, it was terminated with a 4% paraformaldehyde solution.
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