for LDH enzyme assay, after addition of my test sample in the assay reagent, i have recorded decrease in A340nm for approximately 5 mins. i have a data of decrease in OD for 5 mins at interval of 1 min.
how can i calculate ( ΔA340/min test ) ?
Units/ml enzyme= (ΔA340/min test - ΔA340/min blank) *2.7*df / 6.22*0.05
2.7 =total volume of assay
6.22= extinction coefficient of NADH
0.05= volume of sample.
and how to obtain ΔA340/min using the maximum linear rate for both the test and blank?