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firefly2280's Content

There have been 3 items by firefly2280 (Search limited from 23-January 19)


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#179937 S. aureus lysis protocol for LC-MS/MS

Posted by firefly2280 on 04 December 2017 - 03:23 AM in Protein Expression and Purification

Thanks neuron




#179912 How much protein in S. aureus?

Posted by firefly2280 on 22 November 2017 - 06:21 AM in Microbiology

Hi all

 

I want to do a total protein extract from an S. aureus culture but have no idea what density to grow my cells to or how much culture to extract. I need a final conc of 1 mg / ml but want to aviod diluting the extracted protein as much as possible. 

 

Any rough ideas on where to start? 

 

thanks! 




#179907 S. aureus lysis protocol for LC-MS/MS

Posted by firefly2280 on 21 November 2017 - 08:38 AM in Protein Expression and Purification

So sorry this may have been asked before but everyone seems to do something slightly different.

 

We need to extract protein using urea (8M) from S. aureus, avoiding detergent as we will do SDS-PAGE and then LC-MS/MS after. 

 

Ive no idea where to start, need a total of around 1 mg / ml protein but can I just spin down a pellet and add urea and bead beat? Should I add a lysis buffer? 

 

If anyone knows of a rough starting amount of bacteria as well that wil be great but I can work that out if needs be! 

 

thanks

 

 





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