Difference between revisions of "Part:BBa K3788027:Experience"
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how you used this part and how it worked out. | how you used this part and how it worked out. | ||
| − | === | + | ===Immunodetection=== |
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| + | <p>To ensure the localization of our protein we made an immunodetection with anti-GFP antibodies.</p> | ||
| + | <p>We also centrifuged our cells and charged the supernatant in the SDS-PAGE to compare the resultas of the immunodetection with the immunodetection presented in Part:BBa_K3788026.</p> | ||
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| + | https://2021.igem.org/wiki/images/9/93/T--Aix-Marseille--WesternBlot_PelBsfGFP%2BAidaID33-N.png | ||
| + | <p><i><b>Figure :</b> Nitrocellulose membrane showing the results of the immunodetection led on cells and supernatant of bacteria containing pBAD24 and BBa_K3788027 induced or not.</i></p> | ||
| + | <p><i>Controls: Charge control for anti-GFP antibodies.</i></p> | ||
| + | <p><i>Bacteria containing empty pBAD24, induced with 1% arabinose for 5 hours (showed in results of <html><a href="https://parts.igem.org/Part:BBa_K3788026 ">BBa_K3788026</a></html>)</i></p> | ||
| + | <p><i>Rule page Ladder molecular weights (kDa) (lowest to highest band) : 10 ; 15 ; 25 ; 35 ; 55 ; 70 ; 100 ; 130 ; 250</i></p> | ||
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| + | <p>We can’t see any specific band (no bands either at 25kDa or 76 kDa for induced and non-induced conditions). We can’t conclude if our passenger proteins stay linked to the Aida-I autotransporter.</p> | ||
| + | |||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 16:49, 21 October 2021
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how you used this part and how it worked out.
Immunodetection
To ensure the localization of our protein we made an immunodetection with anti-GFP antibodies.
We also centrifuged our cells and charged the supernatant in the SDS-PAGE to compare the resultas of the immunodetection with the immunodetection presented in Part:BBa_K3788026.
Figure : Nitrocellulose membrane showing the results of the immunodetection led on cells and supernatant of bacteria containing pBAD24 and BBa_K3788027 induced or not.
Controls: Charge control for anti-GFP antibodies.
Bacteria containing empty pBAD24, induced with 1% arabinose for 5 hours (showed in results of BBa_K3788026)
Rule page Ladder molecular weights (kDa) (lowest to highest band) : 10 ; 15 ; 25 ; 35 ; 55 ; 70 ; 100 ; 130 ; 250
We can’t see any specific band (no bands either at 25kDa or 76 kDa for induced and non-induced conditions). We can’t conclude if our passenger proteins stay linked to the Aida-I autotransporter.
User Reviews
UNIQc8740338e09b3fb5-partinfo-00000001-QINU UNIQc8740338e09b3fb5-partinfo-00000002-QINU