WESTERN BLOTTING
The bands in the Western Blot were accurate. The transfection of the inserts were verified and assays of tPA were prepared from the lysates.
After having proven the presence of tPA expression in the cells transfected with Western Blotting, the concentration of the amount of tPA in the cell and the amount of secreted tPA were measured using the ‘Human tPA Activity Kit’ to show that the expressed proteins are functionally active.
The real parameter of the measurement in the Assay was the product of the reaction of the plasmin enzyme. Since tPA shifts the inactive plasminogen to active plasmine, the measured value also presents tPA activity. The yield gives absorbance at 405 nm.
tPA ASSAY
File:Https://static.igem.org/mediawiki/2014/0/05/ATOMS-tpa results10.png
According to the tPA assay results we obtained, we were able to prove that the tPA enzyme could be produced and secreted from the cell successfully hence also showing that tPA is functionally active.
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