Application Notes
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IgG purification procedure eample: 1. Wash column with ddH2O to remove air bubbles. 2. fill column with protein L beads. 3. Wash the column with 5X volume of Binding Buffer. 4. Dilute serum sample with Binding Buffer (1:1 ratio). 5. Invert the diluted serum sample to mix well. Make sure no bubbles in the solution. 6. Pour the solution onto the column. 7. Collect the solution and repeat step 6 & 7 for 10 times. 8. Wash the column 5-10 times with the Binding Buffer. 9. Add Elution Buffer to elute IgG. 10. Collect the eluent using microcentrifuge tube. 11. Repeat step 9 & 10 for 10 times. 12. Assay protein concentration and combine the fractions containing sufficient amount of IgG.
Buffer example: Binding buffer: 0.05 M sodium borate, 0.15 M sodium chloride pH 8.0 Elution buffer: 0.1 M citric acid, pH 2.75. Note: Protein L binds to all IgG subclasses from human, mouse and rat species. It also binds to human, mouse, and rat IgM, IgA, IgE, and IgD, as well as Fab and K light chains. Protein L is also superior for binding to chicken, Hamster and pig IgG.
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