Analysis of Host-Cell Responses by Immunoblotting, ELISA, and Real-Time PCR

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 845)

Abstract

Intracellular responses to external pathogens/stimuli are crucial to the host’s response to infection. The methods used to analyse these responses fall into many categories. Activation of proteins as part of a signal cascade can be screened for using conventional immunoblotting techniques or immunoprecipitation to identify the presence of modified proteins or protein complexes. Transcription factor activity can be screened for by EMSA or ELISAs to identify DNA binding of these factors. Finally, expression of activated genes can be quantified using real-time PCR methods. Here, we will show how to perform these assays and discuss the relative merits of each.

Key words

Immunoblotting Transcription factor Cell signalling Immunoprecipitation ELISA Real-time PCR Electrophoresis Phosphorylation Protein modification 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Department of Oral ImmunologyKing’s College London Dental InstituteLondonUK

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