DEPENDS ON THE SUBSTRATE YOU TAKE AND SPECIFIC ACTIVITY OF THE ISOLATED ENZYME
Yes, it will change quite dramatically as you change the substrate. Two common substrates are ABTS and TMB. The latter is 50-100 fold more sensitive. The activity of the enzyme will also be affected by pH, and possibly by the buffer conditions, and therefore there is no meaningful value for working concentration. You should just do serial dilutions from a known concentration of HRP to find the working concentration under your assay conditions.
The actual working concentration of Horseradish peroxidase (HRP) can vary depending on the specific application and experimental conditions. However, commonly used concentrations of HRP in various assays and experiments range from 0.1 to 10 units per milliliter (U/mL).
It's important to note that the activity of HRP is typically measured in units, which represent the amount of enzyme required to convert 1 micromole of substrate per minute under specific conditions. The specific activity of HRP can vary between different sources and preparations, so it's always recommended to consult the manufacturer's instructions or the literature specific to your experimental setup for the recommended working concentration of HRP.
Additionally, the optimal concentration of HRP may also depend on other factors such as the concentration of the substrate, the reaction buffer, and the desired sensitivity of the assay. It's advisable to perform some preliminary experiments to determine the optimal working concentration of HRP for your specific experimental conditions.
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