Difference between revisions of "Part:BBa K3795000:Experience"
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In order to evaluate if DME-C could recognize anti-Dengue antibodies, western blot analysis were performed with two different commercial antibodies: anti-Dengue NS1, and anti-Dengue envelope protein. Fortunately, both DME-C and DME-BR were capable of recognizing anti-Dengue E antibodies, showing that Ammit fullfiled at least its main function. However, the preliminary anti-Zika assays showed that the protein also recognized anti-Zika antibodies. | In order to evaluate if DME-C could recognize anti-Dengue antibodies, western blot analysis were performed with two different commercial antibodies: anti-Dengue NS1, and anti-Dengue envelope protein. Fortunately, both DME-C and DME-BR were capable of recognizing anti-Dengue E antibodies, showing that Ammit fullfiled at least its main function. However, the preliminary anti-Zika assays showed that the protein also recognized anti-Zika antibodies. | ||
| − | https://static.igem.org/mediawiki/parts/9/9f/T--Rio_UFRJ_Brazil--Anti-E-peq-denv.png | + | <br> https://static.igem.org/mediawiki/parts/9/9f/T--Rio_UFRJ_Brazil--Anti-E-peq-denv.png <br> |
| − | <br> Anti-Dengue Envelope (Invitrogen) protein Western blot analysis. We used Anti - Mouse IRDye® 800CW (LI-COR), at 1:10,000, as the secondary antibody. M: Protein marker (Spectra™ Multicolor Low Range Protein, Thermo). 1 - 3: Different dengue virus lysate containing 30 μg of protein used as a positive control. 4: 17 μg of DME-C. 5: 17 μg of DME-BR. | + | Anti-Dengue Envelope (Invitrogen) protein Western blot analysis. We used Anti - Mouse IRDye® 800CW (LI-COR), at 1:10,000, as the secondary antibody. M: Protein marker (Spectra™ Multicolor Low Range Protein, Thermo). 1 - 3: Different dengue virus lysate containing 30 μg of protein used as a positive control. 4: 17 μg of DME-C. 5: 17 μg of DME-BR. |
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| + | <br> https://static.igem.org/mediawiki/parts/2/28/T--Rio_UFRJ_Brazil--Anti-Ns1-peq-Zika.png <br> | ||
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(1)e(2)MW Molecular Weight Marker, Unstained Protein Molecular Weight Marker; (3) 2.0μg DMEC sample; (4) 1.0μg DMEC sample; (5) Sample of 2.0μg DMEC; (6) DMEBR 2.0μg sample; (7) DMEBR 1.0μg sample; (8) DMEBR 0.5μg sample; | (1)e(2)MW Molecular Weight Marker, Unstained Protein Molecular Weight Marker; (3) 2.0μg DMEC sample; (4) 1.0μg DMEC sample; (5) Sample of 2.0μg DMEC; (6) DMEBR 2.0μg sample; (7) DMEBR 1.0μg sample; (8) DMEBR 0.5μg sample; | ||
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Applications of DME-C-BBa_K3795000
In order to evaluate if DME-C could recognize anti-Dengue antibodies, western blot analysis were performed with two different commercial antibodies: anti-Dengue NS1, and anti-Dengue envelope protein. Fortunately, both DME-C and DME-BR were capable of recognizing anti-Dengue E antibodies, showing that Ammit fullfiled at least its main function. However, the preliminary anti-Zika assays showed that the protein also recognized anti-Zika antibodies.
Anti-Dengue Envelope (Invitrogen) protein Western blot analysis. We used Anti - Mouse IRDye® 800CW (LI-COR), at 1:10,000, as the secondary antibody. M: Protein marker (Spectra™ Multicolor Low Range Protein, Thermo). 1 - 3: Different dengue virus lysate containing 30 μg of protein used as a positive control. 4: 17 μg of DME-C. 5: 17 μg of DME-BR.
(1)e(2)MW Molecular Weight Marker, Unstained Protein Molecular Weight Marker; (3) 2.0μg DMEC sample; (4) 1.0μg DMEC sample; (5) Sample of 2.0μg DMEC; (6) DMEBR 2.0μg sample; (7) DMEBR 1.0μg sample; (8) DMEBR 0.5μg sample;
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