What Protein Quantification Methods Are The Best?

[josekolich]

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)

[Ms. DNA]

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.

[daisy]

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.