Table 5.
Protein purification conditions.
| Protein | Lysis buffer | Wash I | Wash II | Elution buffer | Dialysis buffer |
|---|---|---|---|---|---|
| His-MACROD1 | 20 mM Hepes/NaOH, pH 8.0, 300 mM NaCl, 5 mM imidazole, 0.1% NP-40, 10% glycerol, 1 mM TCEP, PIC, Benzonase | 20 mM Hepes/NaOH, pH 8.0, 300 mM NaCl, 10 mM imidazole, 0.1% NP-40 | 20 mM Hepes/NaOH, pH 8.0, 300 mM NaCl, 300 mM imidazole | 30 mM Hepes/NaOH, pH 8.5, 200 mM NaCl, 10% glycerol, 1 mM TCEP | |
| His-MACROD2 | 20 mM Tris–HCl, pH 6.4, 200 mM NaCl, 10% glycerol, 1 mM TCEP | ||||
| His-TARG1 | 20 mM Hepes/NaOH, pH 7.5, 200 mM NaCl, 10% glycerol, 1 mM TCEP | ||||
| His-PARGcat | 20 mM Hepes/NaOH, pH 8.0, 300 mM NaCl, 300 mM imidazole, 1 mM TCEP | 25 mM Hepes/NaOH, pH 8.0, 300 mM NaCl, 10% glycerol, 1 mM TCEP | |||
| His-ARH1 | 50 mM Tris–HCl, pH 8.0, 300 mM NaCl, 5 mM MgCl, 1 mM TCEP, 10 mM imidazole, PIC, Benzonase | 50 mM Tris–HCl, pH 8.0, 150 mM NaCl, 5 mM MgCl, 1 mM TCEP, 10 mM imidazole | 50 mM Tris–HCl, pH 8.0, 150 mM NaCl, 5 mM MgCl, 1 mM TCEP, 300 mM imidazole | 50 mM Tris–HCl, pH 7.0, 150 mM NaCl, 10% glycerol, 1 mM TCEP | |
| His-ARH3 | 50 mM Tris–HCl, pH 7.0, 150 mM NaCl, 10% glycerol, 1 mM TCEP | ||||
| GST-PARP10_ cat | 20 mM Tris–HCl, pH 8.0, 150 mM NaCl, 1 mM EDTA, 5 mM DTT, PIC, Benzonase | 50 mM Tris–HCl, pH 8.0, 150 mM NaCl | 50 mM Tris–HCl, pH 8.0, 150 mM NaCl, 20 mM glutathione | 25 mM Tris–HCl, pH 7.5, 150 mM NaCl, 10% glycerol, 1 mM TCEP | |
| His-mART2.2 | 20 mM Tris–HCl, pH 7.5, 300 mM NaCl, 1 mM TCEP, 500 µg/ml lysozyme, PIC, Benzonase | 50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 1 mM TCEP, 10 mM imidazole | 50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 10% glycerol, 250 mM imidazole | 50 mM Tris–HCl, pH 7.5, 150 mM NaCl, 10% glycerol, 0.5 mM TCEP | |
| His-Pertussis toxin_cat | 25 mM Tris–HCl, pH 8.0, 500 mM NaCl, 10% glycerol, 1 mM TCEP, 10 mM imidazole, PIC, Benzonase | 50 mM Tris–HCl, pH 8.0, 500 mM NaCl, 1 mM TCEP, 10 mM imidazole | 50 mM Tris–HCl, pH 8.0, 200 mM NaCl, 1 mM TCEP, 10 mM imidazole | 25 mM Tris–HCl, pH 8.0, 500 mM NaCl, 300 mM imidazole, 1 mM TCEP | 25 mM Hepes/NaOH, pH 8.0, 500 mM NaCl, 10% glycerol, 1 mM TCEP |
| His-mPARP14- m2m3-Fc | 20 mM Hepes/NaOH, pH 8.0, 300 mM NaCl, 10 mM Imidazole, 0.1% NP-40, 10% glycerol, 1 mM TCEP, PIC, Benzonase | 20 mM Hepes/NaOH, pH 8.0, 300 mM NaCl, 10 mM imidazole, 0.1% NP-40 | 20 mM Hepes/NaOH, pH 8.0, 150 mM NaCl, 10 mM imidazole | 20 mM Tris–HCl, pH 8.0, 300 mM NaCl, 300 mM imidazole, 1 mM TCEP | 25 mM Hepes/NaOH, pH 8.0, 300 mM NaCl, 10% glycerol, 1 mM TCEP |
| His-mPARP14-m2m3-GE-Fc | |||||
| His-DXO | 25 mM Hepes/NaOH, pH 7.0, 300 mM NaCl, 5 mM MgCl, 1 mM TCEP, 10 mM imidazole, 0.1% NP-40, PIC, Benzonase | 25 mM Hepes/NaOH, pH 7.0, 150 mM NaCl, 5 mM MgCl, 1 mM TCEP, 10 mM imidazole | 25 mM Hepes/NaOH, pH 7.0, 200 mM NaCl, 5 mM MgCl, 1 mM TCEP, 300 mM imidazole | 25 mM Hepes/NaOH, pH 7.0, 150 mM NaCl, 10% glycerol, 1 mM TCEP | |