AAT Bioquest

Why is my NADH assay showing unreasonably high absorbance readings?

Posted February 19, 2020


There are many possible reasons for strange and unusable absorbance results, particularly when investigating these enzymatic cofactors. For initial troubleshooting, we recommend:


  • Run a negative control to test instrument settings-photometer settings in particular may vary
  • Check for precipitation, particularly when using tissue homogenizations, as particulate increases scattering
  • Buffer levels too low or unsuitable buffer used-some formulations may cause enzymatic reactions leading to discoloration or frothing. See our buffer recipe and formulation resource in 'Additional Resources' below.
  • If using an in-house procedure, try an NAD(H) assay kit suitable for your needs to investigate if the procedure or the material is the cause
Additional resources

Buffer Preparations and Recipes