Difference between revisions of "Part:BBa K838000:Experience"
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=== Experimental Setup === | === Experimental Setup === | ||
| − | We transfected the CHO (strain dg44) cells the evening before the experiment, turned on the blue light for 2 hours the next morning, took samples and performed the Western blot using [http://2012.igem.org/Team:EPF-Lausanne/Protocol/Western_Blot this] protocol. | + | We transfected the CHO (strain dg44) cells the evening before the experiment with 100% pcDNA3-LovTAP, turned on the blue light for 2 hours the next morning, took samples and performed the Western blot using [http://2012.igem.org/Team:EPF-Lausanne/Protocol/Western_Blot this] protocol. |
We did the same with HEK cells. | We did the same with HEK cells. | ||
Revision as of 00:48, 5 October 2012
Western blot
Idea
LovTAP-VP16 contains the C domain of VP16, therefore with a VP16 antibody it is possible to detect LovTAP in a Western blot.
Experimental Setup
We transfected the CHO (strain dg44) cells the evening before the experiment with 100% pcDNA3-LovTAP, turned on the blue light for 2 hours the next morning, took samples and performed the Western blot using [http://2012.igem.org/Team:EPF-Lausanne/Protocol/Western_Blot this] protocol. We did the same with HEK cells.
Results
Lane 1: VP16 positive control (VP16 protein) Lane 3: Untransfected CHO cells Lane 4-5: LovTAP-VP16 transfected CHO cells Lane 6: Untransfected HEK cells Lane 7-8: LovTAP-VP16 transfected HEK cells.
We can clearly see that the VP16 antibody recognized a VP16 domain in lanes 4-5 and 7-8, which are the LovTAP-VP16 transfected cells. We can thus say that the cells expressed a protein with a VP16 domain.
Flow cytometry
Idea
User Reviews
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