Background-free Detection of Mouse Antibodies on Mouse Tissue by Anti-isotype Secondary Antibodies

J Histochem Cytochem. 2021 Aug;69(8):535-541. doi: 10.1369/00221554211033239. Epub 2021 Jul 20.


Immunodetection on mouse routinely processed tissue via antibodies raised in mice faces cross-reactivity of the secondary anti-mouse reagents with endogenous immunoglobulins, which permeate the tissue. Various solutions to this problem have been devised and include endogenous Ig block with anti-mouse Fab fragments or directly conjugated primary antibodies. Mouse isotype-specific antibodies, differently from reagents directed against both heavy and light chains, fail to detect endogenous Ig after fixation and embedding, while providing a clean and specific detection system for mouse antibodies on mouse routinely processed tissue.

Keywords: antibodies; immunoglobulin isotypes; immunohistochemistry; indicators and reagents; mice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Cross Reactions / immunology
  • Immunoconjugates / chemistry*
  • Immunoglobulin Fab Fragments / chemistry*
  • Immunoglobulin Isotypes / chemistry*
  • Immunohistochemistry / methods*
  • Immunohistochemistry / standards
  • Indicators and Reagents / chemistry*
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / metabolism
  • Lymph Nodes / cytology
  • Lymph Nodes / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Rats
  • Rats, Sprague-Dawley
  • Spleen / cytology
  • Spleen / metabolism


  • Antibodies, Monoclonal
  • Immunoconjugates
  • Immunoglobulin Fab Fragments
  • Immunoglobulin Isotypes
  • Indicators and Reagents