Protein G (Agarose), Fast Flow

Blue Ice

Supplied as 10ml packed beads, containing 2mg/ml Protein G suspended as a 50% slurry in distilled water containing 20% ethanol for a final volume of 20ml. Binding capacity of 20mg human IgG/ml agarose. Prior to use, wash the agarose beads with an appropriate buffer to remove the ethanol.

Stable in all aqueous buffers used in Protein G chromatography. Maximum linear flow rate ≤1300cm/hr. Recommended for flow rates from 30 to 400cm/hr. Useful for purifying IgG from mouse, sheep and rabbit.

Applications:
Suitable for Affinity Purification and Immunoprecipitation. Other applications not tested.

Recommended Dilution:
Antibody Purification: A mouse monoclonal IgG was purified from hybridoma supernatant fraction using a 12ml column of Protein G (Agarose), Fast Flow and a Waters 650E Advanced Protein Purification System.
Elution Profile and Analysis: Mouse IgG was purified from 2L of hybridoma supernatant fraction using Protein G Agarose, Fast Flow. The column eluent was monitored with an UV detector and recorded at 280nm.
Immunoprecipitation: 20ul of a 50% slurry (10ul packed beads) was used to capture a mouse IgG Immunoprecipitation complex. Immunoprecipitation of Phosphotyrosine containing proteins contained in EGFstimulated A431 cell lysate using anti-phosphotyrosine, (4ug) in conjunction with 20ul (50% slurry) Protein G Agarose, fast flow followed by immunoblot analysis using antiphosphotyrosine. Blot was developed using an HRP-conjugated secondary antibody and the ECL detection system.

Storage and Stability:
May be stored at 4°C. For long-term storage, aliquot and store at 4°C. Do not freeze. Aliquots are stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial prior to removing the cap. Further dilutions can be made in assay buffer.