[Protein-analysis] for purification

[jayaraman tharmalingam]

Dear sir,
                i am try to purify the IgG fraction from rabbit serum. i use the proteinA-CL agarose column for best fit to purify the IgG but i encounter some problem with this column.
1) i unable to elute the IgG fraction by 0.1M glycine-Hcl buffer but i try other option with 3M potassium isothiocynate, it elute some reasonable concentration of IgG.
 
2) In ELISA (Antibody capture), i use 3M potassium isothiocynate elute IgG but it not work well.
 
so please somebody help me to resolve my problem.
 
by
jayaraman


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