Altman:Protocols/Protein Purification/SF9 purification
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Small-scale purification of FLAG-tagged proteins from SF9 cells
Prepare the following beforehand:
- LYSIS BUFFER
- Final concentrations:
- 200 mM NaCl
- 4 mM MgCl2
- 20 mM Imidazole, pH 7.5
- 0.5 mM EDTA
- 1 mM EGTA
- 0.5% (v/v) Igepal
- 7% (w/v) sucrose
- 1 mM PMSF (0.25 M stock)
- 10 μg/mL Aprotinin
- 10 μg/mL Leupeptin
- 5 mM DTT
- 2 mM ATP
5 mL total | |
---|---|
2x lysis buffer | 2.5 mL |
1 mg/mL aprotinin stcok | 50 uL |
1 mg/mL leupeptin stock | 50 uL |
1 M DTT | 25 uL |
100 mM ATP | 100 uL |
0.25 M PMSF | 20 uL |
diwater | 2.255 mL |