Unit

UNIT 27.6 Using Antibody Arrays to Measure Protein Abundance and Glycosylation: Considerations for Optimal Performance

  1. Brian B. Haab,
  2. Katie Partyka,
  3. Zheng Cao

Published Online: 24 SEP 2013

DOI: 10.1002/0471140864.ps2706s73

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Haab, B. B., Partyka, K. and Cao, Z. 2013. Using Antibody Arrays to Measure Protein Abundance and Glycosylation: Considerations for Optimal Performance. Current Protocols in Protein Science. 73:27.6:27.6.1–27.6.16.

Author Information

  1. Van Andel Research Institute, Grand Rapids, Michigan

Publication History

  1. Published Online: 24 SEP 2013

Abstract

Antibody arrays provide a valuable method for obtaining multiple protein measurements from small volumes of biological samples. Antibody arrays can be designed to target not only core protein abundances (relative or absolute abundances, depending on the availability of standards for calibration), but also posttranslational modifications, provided antibodies or other affinity reagents are available to detect them. Glycosylation is a common modification that has important and diverse functions in both normal and disease biology. Significant progress has been made in developing methods for measuring glycan levels on multiple specific proteins using antibody arrays and glycan-binding reagents. This unit describes practical approaches for developing, optimizing, and using antibody array assays to determine both protein abundance and glycosylation state. Low-volume arrays can be used to reduce sample consumption, and a new way to improve the binding strength of particular glycan-binding reagents through multimerization is discussed. These methods can be useful for a wide range of biological studies in which glycosylation may change and/or affect protein function. Curr. Protoc. Protein Sci. 73:27.6.1-27.6.16. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • antibody arrays;
  • glycosylation;
  • lectins