Coding

Part:BBa_K3788027:Experience

Designed by: Rebecca Pagčs   Group: iGEM21_Aix-Marseille   (2021-09-30)
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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.


T--Aix-Marseille--WesternBlot_PelBsfGFP%2BAidaID33-N.png

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.


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