Practical Protein Chemistry - A. Darbre 1989

Analytical Methods
Determination of Protein Bound to a Solid Support
Sepharose-Bound Protein

Reagent. Dissolve 50 g of polyethylene glycol (M 20,000) in 50 mL of Water.

Procedure. Place the sepharose-bound protein suspension (0.2 mL – 1 mL) in a quartz cuvette (with a 1 cm optical path length). Add the polyethylene glycol solution with a syringe to a final volume of 3 mL. Mix gently to avoid The formation of air bubbles. Measure the absorbance at 280 nm against a protein-free resin blank.

Plot a linear graph of absorbance versus protein concentration. Sensitivity is 20 µg protein/0.5 mL sepharose. The calibration curve can be constructed using data obtained by adding increasing amounts of protein to the gel suspension (apoglyceraldehyde-3-phosphate dehydrogenase was used).



Last update: 06/08/2026

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