EMC4 Antibody, HRP conjugated

Western Blotting

• Dilution Range: 1:5,000–1:20,000 in WB, validated using lysates from 293, SH-SY5Y, U87-MG, and HepG2 cells .

• Detection: Paired with goat anti-rabbit IgG (H+L) HRP conjugate at 1:1,000 dilution .

Immunohistochemistry

• Protocol: Requires heat-mediated antigen retrieval (EDTA buffer, pH 9) and HRP Polymer for Rabbit IgG .

ELISA

• Sensitivity: Polyclonal HRP conjugates enhance biomarker detection in diagnostic assays by amplifying weak signals .

Conjugation Methodology

HRP conjugation involves site-specific labeling to preserve antibody functionality:

• Traditional Methods: Periodate oxidation of HRP’s carbohydrate moieties, followed by lyophilization and antibody coupling .

• Advanced Protocols: Tools like oYo-Link® HRP enable rapid (<2 hours), site-directed conjugation with 1–2 HRP labels per antibody heavy chain, ensuring consistency and avoiding cross-reactivity .

Comparison of Conjugation Techniques:

Technical Considerations

• Buffer Compatibility: Conjugates tolerate common storage buffers (e.g., PBS with glycerol) .

• Endogenous HRP: Pretreatment with hydrogen peroxide is recommended for IHC to reduce background .

• Stability: Lyophilized HRP-antibody conjugates retain activity for months at -20°C .