Related Experiment Video
Updated: Jul 3, 2026
Enzyme-linked Immunospot Assay (ELISPOT): Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
Published on: November 23, 2010
Protocol to measure the impact of Clostridioides difficile toxins on antibody responses using ELISA, ELISPOT, and
Kaylee M Norman1, Gillian A Lang1, Jimmy D Ballard1
1Department of Microbiology and Immunology, University of Oklahoma Health Sciences, Oklahoma City, OK 73104, USA.
To determine the effects of bacterial toxins on the humoral immune response, accurate measurement of antibody quantity, specificity, and function is critical. Here, we present a murine injection protocol for bacterial toxins and vaccine antigens and collection of plasma via retro-orbital eye bleeds. We describe steps for measuring antigen-specific antibody titers, antibody-secreting plasma cell abundance, and in vitro function of those antibodies. This protocol may be adapted to measure the impact of several bacterial toxins on humoral immunity. For complete details on the use and execution of this protocol, please refer to Norman et al.1.
To determine the effects of bacterial toxins on the humoral immune response, accurate measurement of antibody quantity, specificity, and function is critical. Here, we present a murine injection protocol for bacterial toxins and vaccine antigens and collection of plasma via retro-orbital eye bleeds. We describe steps for measuring antigen-specific antibody titers, antibody-secreting plasma cell abundance, and in vitro function of those antibodies. This protocol may be adapted to measure the impact of several bacterial toxins on humoral immunity. For complete details on the use and execution of this protocol, please refer to Norman et al.1.
