Size-exclusion chromatography is a powerful technique for the size-dependent separation of biomolecules. While standard, non-denaturing mobile phases are often desirable, denaturing mobile phases containing guanidine hydrochloride (or SDS), may be used to insure un-aggregated, monomers for proper size determination. As shown below, denaturation shifts molecules toward larger apparent size and smaller retention volume.
This document discusses the use of size-exclusion chromatography (SEC) for separating biomolecules. It highlights that while standard non-denaturing mobile phases are common, denaturing phases containing guanidine hydrochloride or SDS can be used to ensure un-aggregated monomers for accurate size determination. The text notes that denaturation shifts molecules toward larger apparent sizes and smaller retention volumes.
| Molecular weight range | 4,000-400,000 daltons |
|---|---|
| Column dimensions | 9.4 x 250 mm |
| Column part number | 884973-901 |
| Detection wavelength | 254 nm |
| Flow rate | 2.0 mL / min |
The columns separate biomolecules within a molecular-weight range of 4,000-400,000 daltons.
The column dimensions are 9.4 x 250.
The flow rate is 2.0 mL / min.