Question (76008) | ADP/ATP Ratio Assay Kit (Bioluminescent) (ab65313)

Go to datasheet (ab65313)

Question

We are planning
to purchase your KIT #ab65313 in order to estimate ATP/ADP ratio in
tissue samples.
I have looked at the description of the assay at your web page
(https://www.abcam.com/ps/products/65/ab65313/documents/ab65313%20ADP%20ATP%20Ratio%20Assay%20Kit%20Bioluminescent%20v2%20%28website%29.pdf)
but samples preparation is described here only for cells. I would
greatly appreciate it if you provide me instruction for tissue samples
preparation for this assay.

Answer


We can recommend two protocols:
1. Prepare a single cell suspension from the tissues of interest with any method desired (cells have to stay intact). Then treat the cells as instructed in the protocols section for adherent cells: Remove culture medium/ buffer and treat cells with 50 μl of Nucleotide Releasing Buffer for 5 minutes at room temperature with gentle shaking. Transfer into luminometer plate. (page 8, 4b).
2. Chop the tissues to the finest possible size (keep cool). Then treat cells with 50 μl of Nucleotide Releasing Buffer for 5 minutes at room temperature with gentle shaking. Transfer into luminometer plate.
Please note that the tissue preparation needs to be optimised by the end user since an increased number of dead cells may affect the results.

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