LCHN Antibody, HRP conjugated

Overview of HRP-Conjugated Antibodies

Horseradish peroxidase (HRP)-conjugated antibodies are critical tools in immunoassays (e.g., ELISA, Western blotting) for amplifying signals via enzymatic reactions . These conjugates consist of an antibody chemically linked to HRP, enabling detection through chromogenic, chemiluminescent, or fluorescent substrates .

Conjugation Methods and Optimization

HRP conjugation typically involves crosslinking lysine residues on HRP to antibodies via periodate oxidation or heterobifunctional linkers . A modified periodate method with lyophilization (freeze-drying) enhances conjugation efficiency by preserving activated HRP and enabling higher antibody-to-enzyme ratios :

• Key Steps:

  1. Oxidation of HRP with sodium metaperiodate to generate reactive aldehydes.

  2. Lyophilization to concentrate activated HRP, improving binding capacity.

  3. Conjugation with antibodies at 1:4 molar ratios, followed by stabilization with sodium cyanoborohydride .

Example: IgG Lambda Light Chain HRP-Conjugated Antibody

While "LCHN" is not explicitly defined, the IgG Lambda Light Chain HRP-20758 (Proteintech) serves as a representative HRP-conjugated antibody :

• Host Species: Rabbit IgG

• Applications: Western blotting (1:1,000–1:6,000 dilution).

• Specifications:

  • Molecular Weight: 25 kDa (observed)

  • Storage: -20°C in PBS with 50% glycerol .

Research Advancements

• Recombinant Conjugates: Fusion proteins combining HRP with antibody Fab fragments enable sensitive biosensors, retaining enzymatic and antigen-binding activity .

• Poly-HRP Systems: Multi-enzyme conjugates (e.g., MegaWox™) amplify signals by attaching multiple HRP molecules to a single secondary antibody .

Performance Considerations

• Substrate Compatibility: Chemiluminescent substrates (e.g., Azure Radiance) optimize sensitivity for low-abundance targets .

• Cross-Reactivity: Species-specific secondary antibodies (e.g., goat anti-rabbit HRP) minimize off-target binding .

Protocols and Best Practices

• Buffer Compatibility: Avoid amine-containing buffers (e.g., Tris) during conjugation to prevent interference .

• Validation: Titrate conjugates for optimal signal-to-noise ratios in target applications .