Coding
LovTAP-VP1

Part:BBa_K838000:Experience

Designed by: Nicolas Gobet   Group: iGEM12_EPF-Lausanne   (2012-09-21)
Revision as of 00:50, 5 October 2012 by Mcheraka (Talk | contribs) (→‎Experimental Setup)


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.


WB.JPG

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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