Protein A/G Sepharose Bead
- Immunoprecipitation (IP), Purification (Purif)
- Protein A/G Sepharose can be used for purification of monoclonal and polyclonal antibodies.
- Protein A/G Sepharose is prepared by covalently coupling recombinant Protein A/G (contains five Ig-binding regions of protein A and three Ig-binding regions of protein G, ABIN412503) to 6 % cross-linked sepharose beads. The coupling was optimized to give a high binding capacity for IgG. The capacity of IgG binding could be greater than 10 mg of human IgG per mL of wet gel.
- Bead Ligand
- Protein A,Protein G
- Bead Matrix
- Sepharose beads
- Bead Size
- 90 µm
Binding capacity greater than 20 mg/ml of wet gel
High flow rate
Low falling off of rProtein A/G
pH stability 2-10.
- Reagent Preparation
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
- Assay Procedure
1. Wash column with ddH2O to remove air bubbles.
2. Fill column with protein A/G 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 4 – 5 times with Binding Buffer containing 0.5 M NaCl
9. Wash the column 4 - 5 times with the Binding Buffer.
10. Add Elution Buffer to elute IgG (0.5-1 ml each time).
11. Collect the eluent using microcentrifuge tube.
12. Assay protein concentration and combine the fractions containing sufficient amount of IgG.
13. To regenerate/store column:
a. Wash with 3 volumes of elution buffer.
b. Wash with 3 volumes of distilled water.
c. Store column in 20 % Ethanol/H2O.
- For Research Use only
- Supplied as 50% slurry in 20 % Ethanol/H2O. >1.5mg Protein A/G per ml Sepharose beads
- Handling Advice
- Do not freeze!
- 4 °C
- Expiry Date
- 12 months
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Dong, Nelson, Grayson, Costa, Guidotti: "Clozapine and sulpiride but not haloperidol or olanzapine activate brain DNA demethylation." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, Issue 36, pp. 13614-9, 2008 (PubMed).
- Zhou, Chen, Arora, Hyams, Kozlowski: "Complement C1 esterase inhibitor levels linked to infections and contaminated heparin-associated adverse events." in: PLoS ONE, Vol. 7, Issue 4, pp. e34978, 2012 (PubMed).