Muramic Acid Assay
Categorized: Biochemical Assays
(Purpose to assay muramic acid, a component of peptidoglycan)
Stock Solutions:
- 1.5% (w/v) p-hydroxydiphenyl in 96% ETOH (PHD)
- 1.0 M NaOH
- concentrated H2SO4
- 0.5 M H2SO4
- 4% (w/v) CuSO4-5H2O in H2O
Procedure:
- Bring sample to 0.5 ml with 1.0 M NaOH.
- Incubate at 38oC for 30 min.
- Add 0.5 ml of 0.5 M H2SO4 + 5 ml conc. H2SO4, stopper tube well and place in boiling H2O bath for 5 min.
- Cool and add 0.05 ml CuSO4 + 0.1 ml PHD.
- Stopper tube and incubate at 30oC for 30 min.
- Read absorbance at 560 nm.
Standard curve:
0 – 20 µg muramic acid.
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