Post on 14-Dec-2015
description
Works Done
• Time Course Trypsin Digestion of P135K• Purification of P135K mutant with no AS• Fluoresence of P135K with Hep & AS• Fluoresence of P135K with Hep & no AS• Troubleshooting methods of lysing after overexpression
Future Works• Purification of P135K in AS, buffer exchange to no AS, run iTC• Re-transformation?• Inclusion body refolding?
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Time Course Trypsin Digestion of P135K at 0.5 mg/mL Trypsin
17 kD Band
1. Urea2. 2 M NaCl3. 1.5 M NaCl4. 800 mM NaCl5. 500 mM NaCl6. 250 mM NaCl7. Flow Through8. Supernatant9. Pellet10. PM
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SDS gel of Purification of P135K in no AS
1. PM2. 1.5M NaCl no AS3. 2M NaCl no AS4. 1500 Elute w/ AS5. Fr. Press Sup6. Son + Lysozyme
Sup7. Son + Lysozyme
Pellet8. Sonication Sup9. Sonication Pellet10. Pellet
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SDS gel of Lysing Methods after Overexpression of P135K