BIOCHEMISTRY-DILUTE LDH?

Question

In biochemistry lab, why do you have to dilute the LDH before adding to the assay? (we used E.Coli, and imidazole in lab, I don't know if that is important)

Answer 1

It depends what step you are talking about, though it seems you are referring to after a his-tagged protein purification. To obtain the most accurate estimation of activity or protein concentration, dilution is helpful. We can factor dilutions into any calculation, so we look to simply dilute to a range where we will obtain a meaningful value with assay. If we knew there was going to be a small amount of protein present, we likely would not require dilution. In this case, we know there is a fair amount of sample so dilution is helpful. Feel free to message me for more guided help.