Date of Graduation

12-2013

Document Type

Thesis

Degree Name

Master of Science in Cell & Molecular Biology (MS)

Degree Level

Graduate

Department

Biological Sciences

Advisor/Mentor

Julie Stenken

Committee Member

Ingrid Fritsch

Second Committee Member

Wayne Kuenzel

Keywords

Pure sciences, Biological sciences, Brain, CCL2, Cytokines, IL-6, Immunoassay, Microdialysis

Abstract

In this thesis, different methods to improve the microdialysis collection procedure for cytokines from brain tissue are presented. The first method was based on stopped flow and results indicating that no significant difference in relative recovery between stopped flow and continuous flow are shown. The second method is an antibody bead-based enhancement method. With the antibody bead-based method, a 3.5 fold increase in the collected concentrations of Chemokine (C-C motif) Ligand 2 (CCL2) were observed. However, there was no significant increase in the in vivo collection efficiency of Interleukin-6 (IL-6) using the antibody enhancement. Finally the development of an in-house created bead assay is discussed for the enhanced collection and quantification for molecules that do not have a commercially available bead-based immunoassay kit. The in-house assay when compared to a commercially available kit for CCL2 showed that a successful assay can be made using the technique employed. Using this method a bead based assay could be made for molecules that do not have these commercially available kits.

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