Periodic acid can be used to degrade carbohydrate structures in order to test for the role of glycans in biological assays.
- 15 mL polypropylene tubes
- 50 mM Sodium Acetate Buffer, pH 4.5
- 100 mM Sodium Borate Buffer, pH 8.0
- Sodium Metaperiodate: Sigma cat no. S1878
- Sodium Borohydride: Sigma cat no. 480886
Treatment of soluble protein:
- Bring CPE to ~3 mg/mL in 1 mL PBS
- Add 1 ml 100 mM sodium metaperiodate in PBS (pH 7.2)
- Incubate at 4°C in dark for 18h (O/N)
- Add 1 ml 260 mM glycerol
- Dialyze against PBS
- Protein assay and gel (approx protein conc 1 mg/mL
Treatment of plate bound protein (from reference 2):
- Coat wells with CPE per routine
- Add 0.25 mM sodium acetate buffer, pH 4.5 with 10 mM sodium metaperiodate
- Incubate 2h at 37°C in the dark
- Wash 1X with dH2O
- Add 0.5M sodium borate buffer, pH 8.0, containing 0.5 M Sodium borohydride for 30 min at 25°C
- Use immediately
- Sato C, Inoue S, Matsuda T, and Kitajima K. . pmid:9716421.
- Hino S, Matsubara T, Urisu A, Aoki N, Sato C, Okajima T, Nadano D, and Matsuda T. . pmid:19285013.
- Who has experience with this protocol?
- Email Wayne Shreffler through OpenWetWare