Affinity Chromatography of Rabbit IgG on Protein-A Supports

The specificity of protein A for immunoglobulins of different species and subclasses is given in Table 4.6. 

Rinse a protein A column (4-5 ml bed volume 15 x 22.5 and 15 x 28.5 mm, respectively) with 40 - 50 ml of Soln. A, flow rate 1 ml/min. Dilute 2 - 5 ml of rabbit antiserum with the same volume of Soln. A and apply with the same flow rate to the column. After sample 

Determine the immunoglobulin concentration of the eluate photometrically (cf equation d). Chap. 1.1.7). 

Store the protein solution either in the presence of a biocide as sodium azide or Thimerosal a 4°C or add glycerol to a final concentration of 30-50% (w/v) and store at -20 °C. 

Harlow E, Lane D (1988) Antibodies. A laboratory manual. Cold Spring Harbor Laboratory, p 310 

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