Janet B. Matsen:FLS pathway crude extract LCMS assay

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Steps & Comments

• ?? figure out how to make a table?
• quench
  • acetonitrile --> denature enzymes to stop chemistry
  • pivolate = internal standard

Misc Notes

• pH for buffer chosen base on other metabolite measurement & assay protocols
• PMSF = phenol methylsulfonyl chloride: inhibits things that can break down proteins, supposedly. See Amanda's protein purification book.
• the percise OD you collect cultures at isn't critical.
• she centrifuges before ultracentrifuging because she wanted to compare crude extract (after regular centrifugation) to after ultracentrifugation.
  • ultracentrifugation removes membrane proteins (which are NADH sinks) and desalting might remove formaldehyde sinks like glutathione.