A340 nm assays of ACS and ADH
07 May 2015
Amount of protein to use
- ACS assays:
- ADH assays:
- In SIP:
- 2015/5/13: 45uM ACS worked just enough to detect ADH activity over 30 min.
Amount of NADH to use
- In SIP, ~4mM seems appropriate.
- SIP extracts include FDH and do not run out of NADH.
- See: /Users/jmatsen/Dropbox/In_Progress/CO2_goes_to_methylotrophy/Enzyme_Assays/Nash_SIP3-4/2015_05_13 SIP3-4 ADH assay
Amount of cofactors to use
- Reasonable as of 5/14/2015
- 1 mM CoA, 4 mM ATP, 5mM MgCl2
Tips
- Use of UV-transparent plastic enhances the range at which you can monitor over
- Polystyrene has background absorbance of ~0.17 A340 units on our machine. (source data)
- UV-star plates have a background absorbance of __ on our machine.
Other resources