Hello,

I am doing research on the glycolytic pathway and have been getting unreliable results for my lactate dehydrogenase assays, due to LDH being subject to oxidation.

Does anybody have any relevant knowledge regarding ways in which to limit oxidation during extraction of re-suspended bacterial pellets?

I use glass bead extraction techniques and re-suspend my frozen cells in PIPES or MES buffer . I work on Escherichia coli and Saccharomyces cerevisiae.  

The ratio of cell suspension to glass beads is, one gram of glass beads per mL cell suspension. Raw cell extract is made by centrifugation following glass bead extraction.

We do NMR and absorption spectroscopy assays.

Thanking you in advance.