B-Cell ELISPOT: For the Identification of Antigen-Specific Antibody-Secreting Cells

Hemangi B Shah1, Kristi A Koelsch

  • 1Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA.

The B-cell ELISPOT assay is a sensitive tool that can be utilized to measure total immunoglobulin (Ig) and antigen-specific antibody-secreting cells. Typically, membrane-bound antigen enables binding of antibody secreted by B-cells. Bound antibody is then detected by using an anti-Ig antibody and a colorimetric substrate, resulting in colored spots on the membrane that can be easily enumerated. Here we have described a method to quantitate antigen-specific antibody-secreting cells from the spleen or bone marrow of a vaccinated mouse.