Skip to main content
SpringerLink
Log in

Measuring Enzyme Kinetics of Glycoside Hydrolases Using the 3,5-Dinitrosalicylic Acid Assay

  • Protocol
  • First Online:
Carbohydrate-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2657))

Abstract

Use of the 3,5-dinitrosalicylic acid reagent allows the simple, rapid quantification of reducing sugars. The method can be used for analysis of biological samples or in characterization of enzyme reactions, as new reducing ends are generated when a polysaccharide substrate undergoes hydrolytic cleavage. Presented here is an application of the method in measuring the kinetics of a glycoside hydrolase reaction, including the optimization of the DNSA reagent, and the production of a standard curve of absorbance versus sugar concentration.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Sumner J, Graham VA (1921) Dinitrosalicylic acid: a reagent for the estimation of sugar in normal and diabetic urine. J Biol Chem 47:5–9

    Article  Google Scholar 

  2. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426–428

    Article  CAS  Google Scholar 

  3. Sumner JB, Sisler EB (1944) A simple method for blood sugar. Arch Biochem 4:333–336

    CAS  Google Scholar 

  4. Sumner JB, Noback CV (1924) The estimation of sugar in diabetic urine, using dinitrosalicylic acid. J Biol Chem 62:287–290

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledge support from the Swedish Research Council (Vetenskapsrådet) and the Swedish Energy Agency (Energimyndigheten).

Author information

Authors and Affiliations

  1. Division of Glycoscience, Department of Chemistry, KTH, Royal Institute of Technology, AlbaNova University Centre, Stockholm, Sweden

    He Li & Lauren S. McKee

Authors
  1. He Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lauren S. McKee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lauren S. McKee .

Editor information

Editors and Affiliations

  1. Agriculture and Agri-Food Canada, Lethbridge Research and Development Center, Lethbridge, AB, Canada

    D. Wade Abbott

  2. Department of Biochemistry, The University of British Columbia, Kelowna, BC, Canada

    Wesley F. Zandberg

Rights and permissions

Reprints and permissions

Copyright information

© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Li, H., McKee, L.S. (2023). Measuring Enzyme Kinetics of Glycoside Hydrolases Using the 3,5-Dinitrosalicylic Acid Assay. In: Abbott, D.W., Zandberg, W.F. (eds) Carbohydrate-Protein Interactions. Methods in Molecular Biology, vol 2657. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3151-5_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3151-5_2

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3150-8

  • Online ISBN: 978-1-0716-3151-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation