The in situ detection of enzyme alkaline phosphatase was performed at 3, 7, and 10 days. After removing the culture medium, the wells were washed twice with PBS (Phosphate buffered saline) solution heated to 37 °C. Then, three hundred and twenty milligrams of the Triz reagent (Sigma) were dissolved in 20 mL of deionized water and, added, 7 mg of the Fast Red reagent (Sigma). 2 mL of this solution was discarded and 8 mg of naphthol (Sigma) diluted in 2 mL of dimethylformamide (Merck) were added to form the working solution. One milliliter of this solution was added to each well. The plate was then taken to the incubator in a humidified atmosphere at 37 °C with 5% of CO2 for 30 minutes. The solution was then taken out of the wells, and the plate was allowed to dry at room temperature before being subjected to qualitative and quantitative examination using Image J.
Triz reagent
Manufactured by Merck Group
Triz reagent is a laboratory chemical used in various analytical and research applications. It is a solution formulated to facilitate specific chemical reactions and transformations. The core function of the Triz reagent is to enable targeted chemical processes within controlled laboratory settings. No further details on intended use or application can be provided in an unbiased and factual manner.
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Lab products found in correlation
2 protocols using triz reagent
1
In Situ Alkaline Phosphatase Detection
2
In Situ Alkaline Phosphatase Detection
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Detection of in situ alkaline phosphatase (ALP) was performed at 3, 7 and 10 days. After removal of the culture medium, the wells were washed twice with PBS at 37ºC. Then 320 mg of Triz reagent (Sigma) was dissolved in 20 mL of deionized water, and both 7 mg of Fast Red reagent (Sigma) and 8 mg of Naphthol (Sigma) diluted in 2 mL of dimethylformamide (Merck) were added. Each well received 1 mL of this working solution, and the plate was taken into the incubator in a humidified atmosphere with 5% CO 2 at 37ºC for 30 minutes. After this time, the solution was removed from the wells, and the plate was left to dry at RT for further photographic documentation.
The colored images were converted into binary images using Image J software, and then converted to grayscale, to allow the pixels to be counted for a quantitative analysis.
The colored images were converted into binary images using Image J software, and then converted to grayscale, to allow the pixels to be counted for a quantitative analysis.
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