Shodex Column Recommendation for Membrane Protein Crystallography (with Detergents)

Record: 810

Category: Column Selection
Product: KW-803
Sample: membrane protein

Question
A customer working on membrane protein crystallography (proteins 22–200 kDa, often forming dimers/tetramers) uses detergents such as n-Dodecyl-β-D-Maltoside (DDM), n-Decyl-β-D-Maltoside (DM), n-Octyl-β-D-Glucoside (OG), and n-Dodecyl-N,N-Dimethylamine-N-Oxide (LDAO). Which Shodex SEC column is suitable for this type of analysis?

Answer:
For membrane proteins solubilized in detergents, the recommended column is the Shodex PROTEIN KW-803 (8.0 × 300 mm, 5 µm).

• Separation mode: Aqueous SEC (silica-based, hydrophilic polymer coating).

• MW range: ~5,000–700,000 Da for proteins.

• Recommended conditions: 0.1 M phosphate buffer with ≤0.1% detergent (e.g., SDS, DDM, OG, or LDAO).

This column has been successfully used for membrane protein analysis under mild detergent-containing conditions.
However, detergents can adsorb onto the stationary phase and permanently alter column characteristics, so it is best to dedicate a KW-803 specifically for detergent-based work.

For higher detergent stability or extended lifetime, the LB-806M (polymer-based SEC, 8.0 × 300 mm, 13 µm) is a good alternative. Polymer packings are generally more resistant to surfactant fouling and compatible with light scattering detectors.

See also Shodex application “Standard Proteins (18) – KW-803” for the protein MW calibration range.

Additional Comments

Detergents can adsorb onto or partially remain in the column, permanently altering surface conditions.
For this reason, it’s best to dedicate one column specifically to detergent-containing methods to maintain reproducibility and prevent cross-contamination.

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