Question (55578) | Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

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Question

1. Can I lyse my cells in ALP Assay buffer and use in downstream applications which involve 1) measuring total DNA and 2) using the protein extract for ELISA assays?
2. My cells are growing on a scaffolding of gelatin and hydroyappetite. Will this scaffolding be extracted using the ALP buffer during cell extraction?
3. How much ALP assay buffer should I use per 100 000 cells?
4. What is the recommended homogenization protocol for cells with this kit?

Answer

1) The cells lysed in the ALP assay buffer cannot ideally be used formeasuring DNA and extracting proteins.

2) I am not certain that the assay buffer will digest the scaffolding. Cells may need to be extracted out of scaffold and then processed with the assay buffer.

3 and 4) Use 100 µl of assay buffer with each 10^5 cells with a dounce homogenizer.

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