Mouse-on-Mouse Cross-Reactivity in Tissue Staining

Symptom

High background when using mouse primary antibodies on mouse tissue, caused by secondary antibody binding to endogenous mouse immunoglobulins and Fc receptors rather than only the primary antibody.

1. 1 Secondary antibody binding to endogenous mouse immunoglobulins in tissue
2. 2 Secondary antibody binding to mouse Fc receptors
3. 3 Cross-reactivity between anti-mouse secondary and tissue immunoglobulins
4. 4 Lack of specialized blocking for mouse-on-mouse applications

1. 1 Follow a Mouse-on-Mouse (MOM) staining protocol with specialized blocking reagents
2. 2 Use MOM blocking kit to prevent secondary antibody binding to endogenous mouse IgG
3. 3 Consider using primary antibodies from different species (e.g., rabbit anti-target)
4. 4 Implement Fc receptor blocking step before primary antibody incubation

Related Video (3)

Bilibili (China-Accessible Mirrors) ★ 92

Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)

"Directly demonstrates IHC on paraffin-embedded tissue sections with antigen retrieval and complete workflow, providing context for where mouse-on-mouse cross-reactivity occurs"

Cell Signaling Technology ★ 88

Immunohistochemistry Protocol for Paraffin embedded Tissue Sections

"CST IHC protocol video for paraffin-embedded tissue sections covers the standard technique where secondary antibody interactions with endogenous immunoglobulins manifest as background"

Cell Signaling Technology ★ 76

Better IHC Step 1: Antigen Retrieval

"Focuses on antigen retrieval optimization and troubleshooting fuzzy/high-background IHC staining, directly addressing the symptom of the failure case"