Here is a simple chromatogram of Bovine Serum Albumin (BSA). Yes, BSA is ubiquitous…and this one came from Sigma. The run conditions are detailed in the table.
|Flow Rate [mL/min]||0.800|
|Column Set||P2500 + P3000|
|Column Temp (°C)||30.0|
|Injection volume μL||80.0|
|Mw Weight Average [Da]||65,001|
|Sample dn/dc [mL/g]||0.185|
|Sample IV [dL/g]||0.0311|
The sample shows excellent separation between the monomer and dimer peaks. Calculated molecular weights are within 3% of the expected values and are stable across the width of the peaks showing monodisperse populations are eluting.
Additionally, the 7º (LALS) and 90º (RALS) light scattering measurements overlay demonstrating that at these molecular sizes, only one angle is necessary for highly accurate and reproducible molecular weight measurements.
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