Difference between revisions of "Part:BBa K838000:Experience"

 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
  
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===Applications of BBa_K838000===
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== Western blot ==
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=== Idea ===
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LovTAP-VP16 contains the C domain of VP16, therefore with a VP16 antibody it is possible to detect LovTAP in a Western blot.
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=== Experimental Setup ===
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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.
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We did the same with HEK cells.
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=== Results ===
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Lane 1: VP16 positive control (VP16 protein)
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Lane 3: Untransfected CHO cells
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Lane 4-5: LovTAP-VP16 transfected CHO cells
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Lane 6: Untransfected HEK cells
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Lane 7-8: LovTAP-VP16 transfected HEK cells.
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[[Image:WB.JPG]]
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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.
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== Flow cytometry ==
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=== Idea ===
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===User Reviews===
 
===User Reviews===

Revision as of 19:17, 4 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, 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

UNIQ7d5bedff25bf8d63-partinfo-00000000-QINU UNIQ7d5bedff25bf8d63-partinfo-00000001-QINU