Singh, S. (2025). Optimizing antibody concentrations for immunohistochemistry in neurotrauma research. The Young Researcher, 9(1), 98-133. http://www.theyoungresearcher.com/papers/singh.pdf

Abstract
Objective/research question: What primary and secondary antibody concentrations optimize the visualization of immunohistochemical (IHC) stains for antigens prevalent in neurotrauma research?

Method: IHC was conducted on six different antigens, with one control. A scoring guide was used to objectively determine which IHC stain was visually optimized. Tested concentrations were chosen based on the manufacturer-recommended concentration and concentrations used in past papers.

Results: The antigens and their respective optimized concentrations are as follows: IBA1, P500S250; GFAP, P1000S500; MAP2, P1000S500; AQP4, P50S500; CD83, P50S500; MBP, P1000S250.

Conclusions: Results suggest a gap in the current secondary antibody literature. Future research should use this study as a reference to facilitate neurotrauma research, limit scientific experimentation on animals, and improve data collection. The goal is to ultimately improve TBI pathology comprehension and find therapeutic treatments to ameliorate patient outcomes.

Keywords: Microglia, astrocytes, traumatic brain injury, immunohistochemistry, antibodies, optimization, staining, neuroinflammation, neurotrauma.

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