Shreffler:Periodate Treatment

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Contents

Overview

Periodic acid can be used to degrade carbohydrate structures in order to test for the role of glycans in biological assays.

Materials

Procedure

Treatment of soluble protein:

  1. Bring CPE to ~3 mg/mL in 1 mL PBS
  2. Add 1 ml 100 mM sodium metaperiodate in PBS (pH 7.2)
  3. Incubate at 4°C in dark for 18h (O/N)
  4. Add 1 ml 260 mM glycerol
  5. Dialyze against PBS
  6. Protein assay and gel (approx protein conc 1 mg/mL

Treatment of plate bound protein (from reference 2):

  1. Coat wells with CPE per routine
  2. Add 0.25 mM sodium acetate buffer, pH 4.5 with 10 mM sodium metaperiodate
  3. Incubate 2h at 37°C in the dark
  4. Wash 1X with dH2O
  5. Add 0.5M sodium borate buffer, pH 8.0, containing 0.5 M Sodium borohydride for 30 min at 25°C
  6. Use immediately

Discussion

discuss this protocol

References

  1. Sato C, Inoue S, Matsuda T, and Kitajima K. . pmid:9716421. PubMed HubMed [Sato]
  2. Hino S, Matsubara T, Urisu A, Aoki N, Sato C, Okajima T, Nadano D, and Matsuda T. . pmid:19285013. PubMed HubMed [Hino]
All Medline abstracts: PubMed HubMed


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