Difference between revisions of "Part:BBa M50069:Experience"

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We performed an enzyme-linked immunosorbent assay to determine the functionality and concentration of our device-derived Ara h2 in culture (Figure 4). Because we detected the highest levels of Ara h2 protein in our simple plasmid samples and due to limited number of wells in the ELISA kit we sourced from Elution Technologies, we decided to only analyze this plasmid. We still employed a range of IPTG induction conditions. Our 0mM IPTG sample and the kit-derived 0ng/mL standard acted as our negative controls and the a series of kit-derived standards of known Ara h2 concentration were utilized as the positive controls and reference points.  

We performed an enzyme-linked immunosorbent assay to determine the functionality and concentration of our device-derived Ara h2 in culture (Figure 4). Because we detected the highest levels of Ara h2 protein in our simple plasmid samples and due to limited number of wells in the ELISA kit we sourced from Elution Technologies, we decided to only analyze this plasmid. We still employed a range of IPTG induction conditions. Our 0mM IPTG sample and the kit-derived 0ng/mL standard acted as our negative controls and the a series of kit-derived standards of known Ara h2 concentration were utilized as the positive controls and reference points.  

Our ELISA results enforced the data obtained from our initial Western. We found protein present and able to bind to anti-Ara h2 coated wells at all IPTG induction conditions except for 0mM, at highest concentration in the 0.125mM IPTG induced sample. Our negative controls were working well; absorbance quantified in these wells was not significantly difference from the absorbance measured in our blank well (background). Our positive controls were also successful as measured absorbance in those samples matched what was detailed in the Elution Technologies protocol.  

Our ELISA results enforced the data obtained from our initial Western. We found protein present and able to bind to anti-Ara h2 coated wells at all IPTG induction conditions except for 0mM, at highest concentration in the 0.125mM IPTG induced sample. Our negative controls were working well; absorbance quantified in these wells was not significantly difference from the absorbance measured in our blank well (background). Our positive controls were also successful as measured absorbance in those samples matched what was detailed in the Elution Technologies protocol.  

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