Question (22564) | Alkaline Phosphatase Assay Kit (Colorimetric) (ab83369)

Go to datasheet (ab83369)

Question

Can this kit be used with tissue lysates? If yes, how would the lysates be needed to be prepared - what buffer would be compatible?

Answer

Tissue samples can also be used with this kit and can be treated in a similar manner to cell samples. 
For tissue samples:
- Start with 10-100 mg of the tissue (rinse well in PBS),
- add 500-1000 μl (or ~3-4 volumes) of the assay buffer on ice,
- homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope.
- Spin down the sample and collect the supernatant.
- Load the supernatant unto a 10kda spin column for deproteinzation (if indicated in the data sheet; not required for enzyme assays).
- Use the eluate for your subsequent assays.
- Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve. 

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