I was studying the role of temperature in the sensitivity of creatinine assay and I found in literature that at 37°C it is more sensitive than at 25°C. But when i tried the same assay at both the temperature, the reaction is more sensitive at 25°C than at 37°C. Is it acceptable?
In general the rate of an enzyme reaction will increase with temperature from 25 to 37 degrees so the assay signal should be greater if all other parameters, apart from temperature, are fixed. How are you defining sensitivity? Are both assays operating in the linear range? What is the maximum absorbance value for each assay and signal for the control wells (no enzyme added)? With these answers it may be possible to suggest some areas to look at.
Thanks Nick for the insight. I am doing chemical based assay instead of enzymatic reaction.
My definition of sensitivity is how close are the experimental values to the gold standard values.
Yes, both assays are working in linear range.
Please let me know that in case of chemical based assay, temperature change is having any role to improve the sensitivity of the reaction?
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