Protein Analysis - Animal Models of Acute Neurological Injuries: Injury and Mechanistic Assessments - page 163

Chapter 11
Protein Analysis
Anthony Kian-Fong Liou and Jun Chen
Abstract
In many neurobiological experiments, identifying the presence and/or quantitative change of specifi c
proteins as well as their associating partners will contribute to the understanding of their chronological and
physiological functions in the brain. Methods such as Western blot analysis, electrophoretic mobility shift
assay, immunoprecipitation (IP), etc. are powerful tools when used individually and in combination will
enable the investigator to determine the changes and responses of specifi c proteins in disease or treatment
paradigms. In this chapter, four of the more commonly used protein analytical techniques are presented
here. Nevertheless, with minor modifi cations to the procedures, these methods can be adapted to address
in a wide range of questions in our quest to understand the functions of the brain.
Key words: Western blot, EMSA, IP and chIP
1. Introduction
In neurobiological studies, it is common to use animal models to
verify hypotheses. The various staining techniques available are
very useful but limited to identifying the presence and localization
of specifi c proteins. In this chapter, we present other techniques
and their variations that can be used to ascertain the presence of
specifi c proteins and identifying their associating partner, which
can be another protein or fragments of nucleic acids. The use of
these techniques will add another dimension to the information
we can attained from our animal models and hence expand our
understanding from the results of our studies.
The most widely used technique in ascertaining and quantifying
specifi c protein is the Western blot analysis which is used in con-
junction with an antibody recognizing the protein. This method
was developed in the laboratory of George Stark and named at
W. Neal Burnette nearly 30 years ago ( 1 ). It is an analytical