Kinase Assay - (Mar/10/2014 )
I'm performing a Kinase Assay. The enzyme is made up from a 10 ug to a concentration of 0.1 ug/ml. I need to dilute the enzyme with dilution buffer. I'm given a specific activity plot with the specific activity of the enzyme in nmol/min/mg. How can I determine how much I need to dilute the enzyme based of the specific activity?
you need to determine how many units and a convenient volume of enzyme you want to add to the reaction.
Thank you for your response.
Could you please be a little more specific regarding the determination of units. I have already found the specific activity to be 125 nmol/min/mg and plan on using 10uL of my diluted enzyme. I'm just not sure how much it should be diluted.
how many units do you want in each individual reaction? and what is a convenient volume for you to add?
which kinase are you using?
here is a link to the worthington biochemicals search page, type "kinase" into the search box and you will get a list of kinases with descriptions, references and assays. maybe the information you want is here?