Team:Warsaw/Calendar-Main/13 August 2008

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<var><b>Fig. 1.</b>Protein A detection on the surface of bacteria.<br>
<var><b>Fig. 1.</b>Protein A detection on the surface of bacteria.<br>
1 - <i>E. coli</i> expressing OmpA_Omega_A_Alpha,<br>
1 - <i>E. coli</i> expressing OmpA_Omega_A_Alpha,<br>
-
2 - negative control - <i>E. coli</i> expressing OmpA_Z_Omega,<br>
+
2 - <i>E. coli</i> expressing OmpA_Z_Omega (antibodies bindes to protein Z probably due to similarity of protein A and Z),<br>
3 - <i>E. coli</i> expressing OmpA_A_Alpha,<br>
3 - <i>E. coli</i> expressing OmpA_A_Alpha,<br>
-
4 - negative control - <i>E. coli</i> carrying OmpA_Omega_A_Alpha without inducer,<br>
+
4 - <i>E. coli</i> carrying OmpA_Omega_A_Alpha without inducer,<br>
5 - <i>E. coli</i> expressing OmpA_A_Omega,<br>
5 - <i>E. coli</i> expressing OmpA_A_Omega,<br>
-
6 - negative control - <i>E. coli</i> Top10 without plasmid.</var>
+
6 - <i>E. coli</i> Top10 without plasmid.</var>
<br><br>
<br><br>

Revision as of 18:03, 27 October 2008

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Checking for presence of protein A on the cell membrane

Piotr

  1. Pouring bacteria by drops onto nitrocellulose.
  2. Drying.
  3. Blocking.
  4. Anti-A antibody binding.
  5. Washing.
  6. Anti-rabbit antibody binding.
  7. Developing with BCIP and NBT (see Fig. 1.).

Fig. 1.Protein A detection on the surface of bacteria.
1 - E. coli expressing OmpA_Omega_A_Alpha,
2 - E. coli expressing OmpA_Z_Omega (antibodies bindes to protein Z probably due to similarity of protein A and Z),
3 - E. coli expressing OmpA_A_Alpha,
4 - E. coli carrying OmpA_Omega_A_Alpha without inducer,
5 - E. coli expressing OmpA_A_Omega,
6 - E. coli Top10 without plasmid.

Cloning of alpha to pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_omega

Michał K.

Inoculation of pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_omega plasmids to LB with kanamycin.

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