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2nd week 1st week
Protein G Sepharose High Performance is produced by coupling protein G to highly cross-linked agarose beads by the N-hydroxysuccinimide activation method. Amersham Biosciences recombinant protein G, Mr 17 000, is produced in E. coli and contains two IgG binding regions. The albumin binding region of native protein G has been genetically deleted, thereby avoiding undesirable cross-reactions with albumin.
equilibration Specific binding and wash out Elution Regeneration (equilibration)
Buffers 6 5 4 3 2 1 A Pump B • Columns • Protein G • DEAE UV detector (280 nm) OD280 10 9 8 7 Tube no.
1st week DEAE 2nd week DEAE Protein G 8 4 3 7 5 9 3 7 Serum Milk 2 6 std std 1 2 3 4 5 6 7 8 9 10 _ _ 3rd week Gel slab + +