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.