A: GFP(His)6 (Positivkontrolle)

B: GFP (Negativkontrolle)

C: GFP(His)3

D: GFP(His)4

Säule Packen

1 ml Chitin-Suspension

1 ml Chitin-Suspension

1 ml Chitin-Suspension

1 ml Chitin-Suspension

Säule Waschen

20 ml TBST-CB

20 ml TBST-CB

20 ml TBST-CB

20 ml TBST-CB

Antikörper immobilisieren

3 ml aHis-CBD

3 ml aHis-CBD

3 ml aHis-CBD

3 ml aHis-CBD

Waschen

2 ml TBST-CB

2 ml TBST-CB

2 ml TBST-CB

2 ml TBST-CB

Waschen (Umäquilibrieren)

7 ml MBS-CB

7 ml MBS-CB

7 ml MBS-CB

7 ml MBS-CB

Ligand binden

4 ml GFP-His6 “A”

4 ml GFP “B”

4 ml GFP-His3 “C”

4 ml GFP-His4 “D”

Waschen

25 ml MBS-CB

25 ml MBS-CB

25 ml MBS-CB

25 ml MBS-CB

Vor-Elution

0.5 ml Elutionspuffer
ggfls. wiederholen

0.5 ml Elutionspuffer
ggfls. wiederholen

0.5 ml Elutionspuffer
ggfls. wiederholen

0.5 ml Elutionspuffer
ggfls. wiederholen

Elution

2 x 1 ml Elutionspuffer

2 x 1 ml Elutionspuffer

2 x 1 ml Elutionspuffer

2 x 1 ml Elutionspuffer