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lane 7 OmpA1 + strep after induction<br> | lane 7 OmpA1 + strep after induction<br> | ||
lane 8 OmpA1 + his6 before induction<br> | lane 8 OmpA1 + his6 before induction<br> | ||
lane 9 OmpA1 + strep before induction<br><br> | lane 9 OmpA1 + strep before induction<br><br><br/> | ||
''Note that there were 2 copies of each sample: samples in lanes 2 to 5 were denatured for a 2 minute period as per protocol while samples in lanes 6 to 9 were denatured for a 10 minute period, Sammy S.'' | ''Note that there were 2 copies of each sample: samples in lanes 2 to 5 were denatured for a 2 minute period as per protocol while samples in lanes 6 to 9 were denatured for a 10 minute period, Sammy S.'' | ||
2. Then went on to investigate the appropriate times to induce expression of the OmpA-fusion protein constructs on the E.coli surface using IPTG | 2. Then went on to investigate the appropriate times to induce expression of the OmpA-fusion protein constructs on the E.coli surface using IPTG<br> | ||
'''Week 3 gel'''<br/> | |||
[[Image: 2007-07-06protGel2.jpg|thumb|left|500px]]<br> | |||
'''Gel 2: induced immediately'''<br> | |||
Lane 1: Ladder<br> | |||
Lane 2: Strep No IPTG<br> | |||
Lane 3: Strep denatured after 30 min<br> | |||
Lane 4: Strep denatured after 60 min<br> | |||
Lane 5: Strep denatured after 90 min<br> | |||
Lane 6: Strep denatured after 120 min <br> | |||
Lane 7: His No IPTG<br> | |||
Lane 8: His denatured after 30 min <br> | |||
Lane 9: His denatured after 60 min <br> | |||
Lane 10: His denatured after 90 min <br><br><br><br> | |||
[[Image: 2007-07-06protGel1.jpg|thumb|left|400px]]<br> | |||
'''Gel 1'''<br> | |||
Lane 1: Ladder<br> | |||
Lane 2: His 0 min induction<br> | |||
Lane 3: His 30 min induction<br> | |||
Lane 4: His 60 min induction<br> | |||
Lane 5: His 90 min induction<br> | |||
Lane 6: His 120 min induction<br> | |||
Lane 7: Strep 0 min induction<br> | |||
Lane 8: Strep 30 min induction<br> | |||
Lane 9: Strep 60 min induction<br> | |||
Lane 10: Strep 90 min induction<br> | |||
Lane 11: Strep 120 min induction<br> | |||
Lane 12: His 0 min induction 120 min denature<br> | |||
Revision as of 10:42, 7 July 2007
Stuff done thus far:
1. Started out with assays for the expression of his and strep tags on the E.coli surface. Used denaturation electrophoresis for this a picture is shown below:
6/27 protein denaturation electrophoresis gel
Description: 6/27 denaturation electrophoresis gel
The rightmost lane is the ladder
lane 1 Ladder
lane 2 OmpA1 + strep before induction with IPTG
lane 3 OmpA1 + his6 before induction
lane 4 OmpA1 + strep after induction
lane 5 OmpA1 + his6 after induction
lane 6 OmpA1 + his6 after induction
lane 7 OmpA1 + strep after induction
lane 8 OmpA1 + his6 before induction
lane 9 OmpA1 + strep before induction
Note that there were 2 copies of each sample: samples in lanes 2 to 5 were denatured for a 2 minute period as per protocol while samples in lanes 6 to 9 were denatured for a 10 minute period, Sammy S.
2. Then went on to investigate the appropriate times to induce expression of the OmpA-fusion protein constructs on the E.coli surface using IPTG
Week 3 gel
Gel 2: induced immediately
Lane 1: Ladder
Lane 2: Strep No IPTG
Lane 3: Strep denatured after 30 min
Lane 4: Strep denatured after 60 min
Lane 5: Strep denatured after 90 min
Lane 6: Strep denatured after 120 min
Lane 7: His No IPTG
Lane 8: His denatured after 30 min
Lane 9: His denatured after 60 min
Lane 10: His denatured after 90 min
Gel 1
Lane 1: Ladder
Lane 2: His 0 min induction
Lane 3: His 30 min induction
Lane 4: His 60 min induction
Lane 5: His 90 min induction
Lane 6: His 120 min induction
Lane 7: Strep 0 min induction
Lane 8: Strep 30 min induction
Lane 9: Strep 60 min induction
Lane 10: Strep 90 min induction
Lane 11: Strep 120 min induction
Lane 12: His 0 min induction 120 min denature
