Purification and Detection of Etx Pore

Chemicals were purchased from Sigma, UK, unless otherwise stated. The polyclonal antibody against Etx mutant Y30A-Y196A was raised in rabbits as described in ref. ^{51 (link)}. After electrophoretic separation purified Etx pore was detected by SimplyBlue staining (Invitrogen Ltd, Paisley, UK). Protein concentrations were determined using the Pierce™ BCA Protein Assay Kit (Fisher Scientific Ltd, Loughborough, UK). Synthetic oligonucleotides were sourced from Eurofins Genomics, Germany.

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Savva C.G., Clark A.R., Naylor C.E., Popoff M.R., Moss D.S., Basak A.K., Titball R.W, & Bokori-Brown M. (2019). The pore structure of Clostridium perfringens epsilon toxin. Nature Communications, 10, 2641.

Publication 2019

Pierce™ BCA Protein Assay Kit Synthetic oligonucleotides

Antibody Assay Electrophoretic Oligonucleotides Protein Rabbits

Corresponding Organization :

Other organizations : University of Leicester, University of Wolverhampton, Institut Pasteur, Birkbeck, University of London, University of Exeter

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Variable analysis

independent variables

Etx mutant Y30A-Y196A

dependent variables

Etx pore formation
Protein concentrations

control variables

Chemicals purchased from Sigma, UK, unless otherwise stated
Synthetic oligonucleotides sourced from Eurofins Genomics, Germany

controls

Positive control: None mentioned
Negative control: None mentioned

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