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• Protein Extraction from Candida albicans

Protein Extraction from Candida albicans

Instrument: Precellys® 24

Sample: Candida albicans

Protocol:

• Add C. albicans cells to the 2 ml tough microorganism lysing kit.
• Add lysis buffer*: Tris-HCl, pH 5, glycerol, TritonX100, SDS, NaCl, NaF, Sodium orthovanadate, glycerol phosphate, sodium pyrophosphate, EDTA, PMSF with 1x protease inhibitor cocktail (Sigma).
• Run the Precellys at 5000 rpm, 4 x 30 sec with 30 sec breaks on ice.
• Centrifuge at 13,000 rpm at 4°C for 10 minutes.

*This lab was looking to extract and analyze proteins.  You may substitute other buffers if they are more appropriate for your application.

Results:

The study investigated MAP kinase activation in C. albicans in the presence of Streptococcus bacteria.  Homogenization with the Precellys enabled the lab to see clear changes in protein expression.

Western immunoblot analysis of effects of 10mM H₂O₂ on phosphorylation of C. albicans MAP kinases after 20 min in the presence or absence of S. gordonii