I would be grateful if anyone can help clarify laemmli buffer dilution for me.. I needed less dilution of my protein samples with Laemmli buffer (i usually use 2x Laemmli buffer), so I decided to use 4x laemmli buffer (recipe from molecularstation.com webpage-i cannot post the URL add-new user) the ratio i used was 3:1 (Protein sample:buffer) . However, I noted that the sample seem less blue than the samples receiving 2x Laemmli buffer(is this expected?)
and the recipe mentioned that the concentration of bromophenol to be used is 0.01%, but the final concentration is to be 0.02%. How is this possible?