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Host-Fungus Interactions

Volume 845 of the series Methods in Molecular Biology pp 345-360

Date:

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

  • David L. MoyesAffiliated withDepartment of Oral Immunology, King’s College London Dental Institute
  • , Julian R. NaglikAffiliated withDepartment of Oral Immunology, King’s College London Dental Institute Email author 

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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