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Calorimetric assay alp assay

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The Calorimetric assay ALP assay is a laboratory equipment used to measure the activity of the enzyme alkaline phosphatase (ALP) in a sample. The assay relies on the conversion of a colorless substrate into a colored product, which is then quantified using a spectrophotometer. The core function of this assay is to provide a quantitative assessment of ALP levels in various biological samples, such as serum, cell lysates, or tissue homogenates.

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Lab products found in correlation

Picogreen assay, by Thermo Fisher Scientific (2 mentions) Adipo red staining assay, by Lonza (2 mentions) Fast blue rr, by Merck Group (2 mentions)

2 protocols using calorimetric assay alp assay

1

Evaluation of Mesenchymal Stem Cell Differentiation

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hMSCs were cultured for 14 days in differentiation induction medium prior to assessing differentiation. To analyze adipogenic differentiation, Adipo Red staining assay for staining intracellular triglycerides was performed on fixed cells as per manufacturer’s protocol (Lonza). For analyzing osteogenic differentiation, cells were either fixed or lysed at day 14. Alkaline phosphatase (ALP) was assessed by staining fixed cells using Fast Blue RR (Sigma) or by performing calorimetric assay ALP assay (Biovision Inc) on cell lysate as per manufacturer’s protocol. DNA content in cell lysate was determined by performing the PicoGreen assay (Life Technologies) as per manufacturer’s protocol. Analysis of ALP activity using calorimetric ALP assay was performed and represented after normalization by DNA content.
Dhaliwal A., Pelka S., Gray D.S, & Moghe P.V. (2018). Engineering Lineage Potency and Plasticity of Stem Cells using Epigenetic Molecules. Scientific Reports, 8, 16289.
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2

Assessing hMSC Differentiation

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hMSCs were cultured for 14 days in different substrates in induction medium prior to assessing differentiation. To analyze adipogenic differentiation, Adipo Red staining assay for staining intracellular triglycerides was performed on fixed cells as per manufacturer’s protocol (Lonza). For analyzing osteogenic differentiation, cells were either fixed or lysed at day 14. Alkaline phosphatase (ALP) was assessed by staining fixed cells using Fast Blue RR (Sigma) or by performing calorimetric assay ALP assay (Biovision Inc) on cell lysate as per manufacturer’s protocol. DNA content in cell lysate was determined by performing the Pico green assay (Life Technologies) as per manufacturer’s protocol. Analysis of ALP activity using calorimetric ALP assay was performed and represented after normalization by DNA content.
Dhaliwal A., Brenner M., Wolujewicz P., Zhang Z., Mao Y., Batish M., Kohn J, & Moghe P.V. (2016). Profiling Stem Cell States in Three-Dimensional Biomaterial Niches using High Content Image Informatics. Acta biomaterialia, 45, 98-109.
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