Will an ACQUITY Protein Premier SEC 250A column separate enzymes (proteins) with molecular weights of 30 kDa, 40 kDa, and 50 kDa from each other? - WKB236407

ENVIRONMENT

• UltraPerformance Liquid Chromatography (UPLC)
• Ultra High Performance Liquid Chromatography (UHPLC)
• High Performance Liquid Chromatography (HPLC)
• ACQUITY Premier Protein SEC 250A
• Size exclusion chromatography (SEC)
• MaxPeak Premier
• High Performance Surfaces (HPS)

ANSWER

No

ADDITIONAL INFORMATION

Typically, in order to separate one enzyme (protein) from another in SEC when all enzymes have a similar structure in solution, there needs to be at least a two-fold difference in molecular weight for baseline resolution.

For example, you should be able to separate a 30 kDa enzyme from a 60 kDa enzyme, but not a 30 kDa enzyme from a 40 kDa or 50 kDa enzyme.

Also note that analytes are separated by their hydrodynamic radius (size in solution), as opposed to their molecular weight. As a result, use of molecular weight is only a general guide, but true size in solution would be determined based on experimentation.