Team:Carnegie Mellon/Protocols
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His-Tag Soluble Protein Extraction
- Centrifuge 1.5 ml of culture for 1min full speed at 15000 rpm.
- Remove supernatant.
- Add 250 µl of extraction buffer (1% octyl-beta-thioglucoside in 10mM Tris-Cl, pH7.5).
- Incubate at room temperature for 10 min.
- Remove 50 µl of Ni-NTA in 10 mM Trischloride bead solution and place in new tube for each purification.
- Add 1 mL buffer (see table).
- Centrifuge at 800 rpm for 30 seconds.
- Remove supernatant.
- Repeat 3 times and resuspend in 1 mL of Bead Wash Solution.
Estrogen Sensor
Estrogen Sensor Protocol