WTA preparations were separated on tricine polyacrylamide gels using a Bio-Rad tetra cell according to a previously described method (Brignoli et al., 2022 (link)). The gels were separated at 4°C using a constant amperage of 40 mA under constant stirring until the dye front reached the bottom. Gels were washed three times in MilliQ H2O followed by staining in 1 mg/ml Alcian blue overnight. Gels were subsequently destained in in MilliQ H2O, until the WTA became visible and finally imaged.
Douglas E.J., Palk N., Brignoli T., Altwiley D., Boura M., Laabei M., Recker M., Cheung G.Y., Liu R., Hsieh R.C., Otto M., O'Brien E., McLoughlin R.M, & Massey R.C. (2023). Extensive remodelling of the cell wall during the development of Staphylococcus aureus bacteraemia. eLife, 12, RP87026.
Other organizations :
University of Bristol, University of Bath, University of Milan, University of Tübingen, University of Exeter, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Trinity College Dublin, APC Microbiome Institute, University College Cork
Separation method: Tricine polyacrylamide gel electrophoresis (Tricine-PAGE)
dependent variables
Visibility of WTA on the gel after staining and destaining
control variables
Temperature: 4°C
Amperage: Constant 40 mA
Stirring: Constant stirring
Staining: 1 mg/ml Alcian blue overnight
controls
Positive control: Not mentioned
Negative control: Not mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to
get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
