What is required to calibrate size exclusion chromatography?

To effectively calibrate size exclusion chromatography, it is essential to utilize proteins with known molecular weights. Size exclusion chromatography separates molecules based on their size; thus, establishing a relationship between the elution volume or time and the size of the molecules is crucial for accurate analysis. By using proteins with well-defined molecular weights, a standard curve can be created, allowing for the determination of the sizes of unknown proteins when they elute from the column.

The known molecular weights serve as references that enable the calibration of the chromatographic system, resulting in a reliable estimation of the sizes of other proteins being analyzed in subsequent runs. This is fundamental for interpreting the results and understanding the behavior of various biomolecules during the chromatography process. Using other parameters, such as charge, binding affinities, or hydrophobicity, does not directly relate to the calibration of size exclusion chromatography, which specifically focuses on molecular size.