Looking for a good methodology for the protein extraction from porcine oral saliva for MALDI-tof analysis in bacterial adherence studies?

22 June 2015 1 9K Report

I wanted to experience a good methodology for the protein extraction from porcine saliva for MALDI-of analysis of bacterial adherence studies.

I am aware of using the solubilization buffer ( 7M Urea, 2M thiourea, 1% (w/v) CHAPS and I% Triton x-100, 1% ampholytes (3-10) and 1mM TCEP in the proportion of 2:1 of saliva) but don’t know effect of reagents used above on the protein MALDI-tof analysis?

Ulla-Maja Bailey

Hi Asim,

Methods for isolating proteins from human saliva in the following publications may be helpful:

Y. Xu, U. M. Bailey, C. Punyadeera and B. L. Schulz. Identification of salivary N-glycoprotein and measurement of glycosylation site occupancy by boronate glycoprotein enrichment and liquid chromatography/electrospray ionization tandem mass spectrometry. Rapid Commun Mass Spectrom. 2014. 28(5):471-82.

U. M. Bailey, C. Punyadeera, J. Cooper-White and B. L. Schulz. Analysis of the extreme diversity of salivary alpha-amylase isoforms generated by physiological proteolysis using liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed life Sci. 2012. 911:21-6

Good luck

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