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Chemical degradation ~Smith degradation

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Introduction Protocol References Credit lines
Category
Isolation & structural analysis of glycans
Protocol Name

Chemical degradation ~Smith degradation

Authors
 Shunji Natsuka *
Department of biology, Faculty of science, Niigata University

Shin-ichi Nakakita
Department of Functional Glycomics, Life Science Research Center, Kagawa University
*To whom correspondence should be addressed.
KeyWords
Reagents
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sodium metaperiodate (Wako Pure Chemicals, Osaka, Japan)

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sodium acetate (Wako Pure Chemicals, Osaka, Japan)

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sodium borohydride (Wako Pure Chemicals, Osaka, Japan)

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acetic acid (Wako Pure Chemicals, Osaka, Japan)

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ammonia solution (Wako Pure Chemicals, Osaka, Japan)

Methods
1.

Smith degradation

1) 

 Partially oxidize a pyridylamino (PA-) sugar chain (0.01 – 1 nmol) with a mixture of 20 μL of 50 mM sodium metaperiodate solution and 20 μL of 0.1 M sodium acetate, pH 4.0 at 0°C for 15 min in the dark.

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2) 

 Add 40 μL of 0.26 M sodium borohydride solution.

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3) 

 Incubate at 25°C for 1 h.

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4) 

 Add 10 μL of acetic acid and mix well.

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5) 

 Neutralize with aqueous ammonia solution.

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6) 

 Dry up.

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7) 

 Analyze the cleaved PA-sugar chain by HPLC as described in the other protocol (Fluorescent labeling of glycans and HPLC analysis).

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Initial amount

>10 pmol

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Date of registration：2012-11-27 13:46:12
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