Protein A Sepharose CL-4B Column, 5ml, 1 column

Ligand Recombinant Staphylococcus Protein A (rSPA) expressed in E. coli, which contains all five immunoglobin binding domains (E,D,A,B,C) and has a molecular weight of 46.6 kDa.

Bead structure 4 % cross-linked agarose

Dynamic binding capacity >30 mg human IgG/ml medium

pH stability Long term: 3–9

Short term: 2–10

Chemical stability All commonly used aqueous buffers, 6 M guanidine-hydrochloride, 8 M urea.

Physical stability Negligible volume variation due to changes in pH or ionic strength.

Binding buffers 20 mM sodium phosphate, pH 7.0 or 50 mM Tris-HCl buffer, pH 7.0 is recommended as a general method.

Elution buffers 0.1 M glycine buffer pH 3.0 or 0.1 M citric acid pH 3.0 is recommended as a general method. To elute very strongly binding IgGs it may be necessary to lower the pH below 3.0.

We recommend adding appropriate volume of 1M Tris-HCl, pH 9.0 to neutralize the eluted fractions as a safety measure to preserve the activity of acid labile IgGs.

Regeneration Washing the column with 2 to 3 bed volumes of elution buffer.

Cleaning-In-Place Washing the column with 2 column volumes of 6 M guanidine hydrochloride. Immediately re-equilibrate with at least 5 column volumes of binding buffer.

Remove strongly bound hydrophobic proteins, lipoproteins and lipids by washing the column with a non-ionic detergent (e.g. 0.1% Triton™ X-100) at 37°C for one minute. Immediately re-equilibrate with at least 5 column volumes of binding buffer.

Sanitization Sanitize the packed column with 2% Hibitane and 20% ethanol for 6 hours or with 70% ethanol for 12 hours.

Temperature Stability 2-40 °C

Storage Medium should be stored in neutral pH at 2°C to 8°C in the presence of a bacteriostat, e.g. 20% ethanol. Medium must NOT be frozen.

Background Protein A is a cell wall component produced by several strains of Staphylococcus aureus that consists of a single polypeptide chain and contains little or no carbohydrate. Recombinant Protein A is produced in E. coli and functions essentially the same as native Protein A. It consists of 5 IgG-binding domains E-D-A-B-C aligned in series.