For research use only!
Shipping: shipped at ambient temperature
Storage Conditions: store at 4 °C
do not freeze
Shelf Life: 12 months after date of delivery
Form: supplied as a suspension in 20 % ethanol
Protein G resin allows batch or column purification of classes, subclasses and fragments of immunoglobulins from cell culture media and biological fluids. Recombinant Protein G is covalently bound to the matrix, avoiding protein loss and allowing re-use of the resin.
Degree of substitution: 3 mg Protein G / ml resin
Binding capacity: 20 mg human IgG / ml resin
Recombinant Protein G contains only highly homologous IgG-binding domains. The binding site is located on the Fc region of immunoglobulin. In the native Protein G include albumin-binding domain, as well as cell wall and cell membrane binding domains, have been removed to ensure the maximum specific IgG binding capacity. Protein G has affinity for IgG from a variety of mammalian species and for some IgA and IgM.
1. Removal of the storage solution
Wash the beads with 5 - 10 column volumes of distilled water to remove the storage solution.
Please note: For batch purification, remove the storage solution by washing the product on a medium porosity sintered glass funnel.
2. Equilibration of the Protein G Agarose resin
Equilibrate the column with 5 - 10 column volumes of binding buffer.
Binding buffer: IgG from most species binds at neutral pH. The buffers used most frequently are sodium phosphate (25 mM) pH 7.0.
3. Application of the sample
Once the resin is equilibrated, the sample containing the immunoglobulin for purification is applied.
4. Washing of the Protein G Agarose resin
Wash with the binding buffer until OD280 reaches baseline level.
5. Elution of the pure immunoglobulin
Elution is usually achieved at reduced pH. Depending on the sample it may be necessary to decrease pH below 3.0. Most immunoglobulins are eluted in glycine (100 mM) or citric acid buffer (100 mM) at pH 2.5 - 3.0.
Please note: It is recommended to add 0.15 ml of a buffer at pH 9.0 (e.g 1 M Tris) per ml of purified immunoglobulin to neutralize the eluted fractions.
The following procedure describes how a column is packed from bulk affinity resins:
Maximum flow rate: ≥500 cm/h1
Maximum pressure: 21.8 psi (1.5 bar)1
1 at 15 cm bed height