I would like to use above kinase assay kit to check the activity of ATPase in my expressed protein. But I couldn't find a person who has used it in my department. I would like if any body who has used it before can share the experience with me.
1. Calibration series preparation
2. 1X kinase assay reagent preparation
are not clear in protocol.
If you have any videos please share with me.
Thank you.
There protocol is straightforward. You should perform the calibration series with your ATP and ADP stocks before doing your actual experiment. For kinase assays it works very well. The 1mM max limitation was an issue for some ATPases but its still workable.
Hi Nilmini, I have not used this particular kit, but the protocol which u uploaded seems very straight forward to me too. Do you have any specific concerns regarding the calibration series?
About the 1x assay/detection reagent preparation- if you have a 400 assay kit, it is already in solution form and for larger size kits there is a lyophilized powder which needs to be dissolved in the buffer as described in detail (it is a common approach with Promega and then they give a single protocol sheet for all sizes).
Good Luck.
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