Transfection 1: Transfecting TRE:Syk-TEVp for Western Blot to compare the levels of endogenous vs exogenous Syk and to compare levels of expression under different amounts of dox induction. Concentrations of DNA: | Dummy | 1230 ng/ul | | hEF1a:eYFP | 1045 ng/ul | | hEF1a:rtTa | 169.4 ng/ul | | TRE:Syk-TEVp | 871 ng/ul |
Volumes of DNA added: | Well 3 | Well 4 | Wells 7-12 | | Dummy 0.81 ul | Dummy 0.49 ul hEF1a:eYFP 0.49 | hEF1a:eYFP 0.38 ul hEF1a:rtTA 1.18 ul TRE:Syk-TEVp 0.46 ul |
Well 1 HEK293 | Well 2 EMPTY (Would have been B cells) | Well 3 HEK293 Dummy DNA 1000 ng | Well 4 HEK293 hEF1a:eYFP 400 ng Dummy DNA 600 ng | Well 5 EMPTY | Well 6 EMPTY | Well 7 HEK293 hEF1a:eYFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 0 nM Dox | Well 8 HEK293 hEF1a:eYFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 1 nM Dox | Well 9 HEK293 hEF1a:eYFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 10 nM Dox | Well 10 HEK293 hEF1a:eYFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 100 nM Dox | Well 11 HEK293 hEF1a:eYFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 1000 nM Dox | Well 12 HEK293 hEF1a:eYFP 400 ng hEF1a: rtTA 200 ng TRE: Syk-TEVp 400 ng 2000 nM Dox | | | | | | | | | | | | | | |
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