TransfectionWell 1 HEK293 | Well 2 B-Cells | Well 3 HEK293 Dummy DNA 1000 ng | Well 4 HEK293 hEF1a: YFP 400 ng Dummy DNA 600 ng | Well 5 EMPTY | Well 6 EMPTY | Well 7 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 0 nM Dox | Well 8 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 1 nM Dox | Well 9 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 10 nM Dox | Well 10 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 100 nM Dox | Well 11 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 1000 nM Dox | Well 12 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 2000 nM Dox |
Gels:
We have the any kD gels in the lab (expire 2015) and this will work for both our western blot and antibody group's western blot.
If we run out of the above gel we are considering getting the 4-25% gel in order to optimize our resolution. This will also work for both our western blot and antibody group's western blot. The gels currently in the lab have 10 lanes. Here is the plan for the gel: | Lane 1 | Lane 2 | Lane 3 | Lane 4 | Lane 5 | Lane 6 | Lane 7 | Lane 8 | Lane 9 | Lane 10 |
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| Ladder | Well 2 B-cells | Well 3 HEK293 Dummy DNA 1000 ng | Well 4 HEK293 hEF1a: YFP 400 ng Dummy DNA 600 ng | Well 7 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 0 nM Dox | Well 8 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 1 nM Dox | Well 9 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 10 nM Dox | Well 10 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 100 nM Dox | Well 11 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 1000 nM Dox | Well 12 HEK293 hEF1a: YFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 2000 nM Dox |
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