HDPCs were cultured with 1α,25(OH)2D3 for 7 days. For ALP staining, media were removed and the cells were xed with 70% ethanol for one hour. After rinsing with deionized water three times, 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro-blue tetrazolium (NBT) solution (Sigma- Aldrich) was added to each well. The reaction was stopped by the addition of water. The stain was photographed under a HP Officejet Pro L7580 scanner. For quantitative analysis, the stain was extracted with 10% (w/v) cetylpyridinium chloride (Sigma) for 15 min. The stain intensity was quantified by measuring the absorbance at 570 nm on an absorbance microplate reader (Bio-Tek Instruments).
5 bromo 4 chloro 3 indolyl phosphate bcip nitro blue tetrazolium solution
Manufactured by Merck Group
Sourced in United States
5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro-blue tetrazolium (NBT) solution is a chromogenic substrate used for the detection of alkaline phosphatase activity in various biochemical and molecular biology applications. It provides a blue-purple colored precipitate upon enzymatic cleavage.
Automatically generated - may contain errors
2 protocols using 5 bromo 4 chloro 3 indolyl phosphate bcip nitro blue tetrazolium solution
1
Alkaline Phosphatase Staining in HDPCs
2
Zebrafish Embryo in situ Hybridization
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Whole-mount in situ hybridization was carried out as described previously45 (link). Antisense RNA probes for alkbh3 mRNA were labeled with digoxigenin (Roche, Switzerland). Zebrafish embryos at different stages were fixed in 4% paraformaldehyde and incubated with digoxigenin-labeled RNA probes. Alkaline phosphatase-coupled anti-digoxigenin antibody (Roche, Switzerland) was used to detect hybridized probes, and 5-bromo-4-chloro-3-indolyl phosphate (BCIP)/nitro blue tetrazolium (NBT) solution (Sigma, USA) was used as the chromogenic substrate.
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