josekolich Posted May 7, 2004 Share Posted May 7, 2004 What protein quantification methods can be employed with enzimes? What are the basis and methodologies to be used for each case?(Please explain it to me as much as possible) Link to comment Share on other sites More sharing options...
Ms. DNA Posted May 7, 2004 Share Posted May 7, 2004 I've never used enzymes to quantify proteins. Instead I use colormetric assays that use stains such as Bradford Reagent or the BCA protein assay. Link to comment Share on other sites More sharing options...
daisy Posted May 28, 2004 Share Posted May 28, 2004 Well, enzymes ARE proteins!!! After you determine a standard curve using Bradford or Lowry assays, you do a Western Blot; where you separate your proteins on a gel, transfer the proteins to a membrane which you then incubate with an antibody directed against your protein of interest. If you run a loading control of known concentration to check that you've loaded the same amount of protein per well, you can quantify your protein of interest. This of course is a hugely condensed version but look up Sambrook and Maniatis, the Bible, and it will guide you through it. I can send you more detailed instructions if you want via e-mail. Link to comment Share on other sites More sharing options...
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