Protocols (Materials and Methods)
Chitin Content Assay 1
 Wang, Z. et al., Infection and Immunity, 67(1999) 6619-6630.
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Materials
Protocol
Results
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  Principle and General Applications
       Chitin content is measured by a modification of the procedure described by Yabe et al. J. Bacteriol. 178 (1996) 477-483.
 

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  Materials
  Elson-Morgan Reagent
Acetyl Acetone Reagent
1.06 g Na2CO3 and 20 ml H2O and 0.4 ml acetyle acetone
Ehrlich Reagent
534 mg p-dimethylaminobenzaldehyde (P-DMAB) and 10 ml ethanol and 10 ml HCl
 

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  Protocol
  1. Harvest log-phase yeast cells from 20-ml cultures.

2. Suspend in 4 M HCl (1 ml) and boil for 4 hr.

3. Dilute hydrolysates with H2O (19 ml).   Determine by the Elson-Morgan method (Boas, 1953) the amount of hexosamine in a dilutent (1 ml), using N-acetyl-D-glucosamine (Sigma, St. Louis, MO) as a standard. 

4. Determine cell dry weights for calculation of chitin content per milligram of cells by collecting the cells from 20-ml cultures on preweighed 0.45 µm membrane filters (Type HA, Millipore Corp., Bedford, MA).  Dry the sample at 65°C to a constant weight.
 

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  Results
      
 

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  Tips
      
 

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