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) ★ 80

Immunohistochemistry on Paraffin-Embedded Sections (CST Demo)

"Core IHC protocol on paraffin sections, showing the context where MOM cross-reactivity occurs and proper staining steps."

Bilibili (China-Accessible Mirrors) ★ 75

Immunohistochemistry Experiment (IHC) Protocol

"Hands-on IHC protocol demonstrates blocking and staining steps critical for reducing endogenous IgG background."

Cell Signaling Technology ★ 70

Immunohistochemistry Protocol for Paraffin embedded Tissue Sections

"Official IHC protocol video includes troubleshooting tips for high background issues like mouse-on-mouse staining."