This conjugate is primarily used for detecting LIFR expression in:
ELISA: Quantitative analysis of LIFR in serum or cell lysates .
Western Blotting: Identification of LIFR protein in denaturing conditions .
Immunohistochemistry (IHC): Localization of LIFR in tissue sections (though less common due to HRP limitations) .
Antigen Capture: LIFR protein binds to immobilized primary antibody.
Signal Amplification: HRP-conjugated secondary antibody binds, oxidizing substrates (e.g., TMB) to generate measurable products .
Advantages: Homogeneous conjugates with defined stoichiometry (e.g., 1:2 antibody-to-HRP ratio) .
Challenges: Ensuring functional activity of both HRP and antibody domains .
| Parameter | Method/Standard | Source |
|---|---|---|
| Purity | Protein G affinity chromatography (>95%) | |
| Specificity | ELISA, western blot validation | |
| Conjugate Stability | SEC (Size Exclusion Chromatography) |
In studies using HRP-conjugated goat anti-human IgG, antibodies like 1G11 blocked LIF binding to LIFR but not gp130, confirming receptor-specific interactions .
Antibody Titers: Serum antibodies against LIFR achieved titers up to 1:3,200 in ELISA, validating epitope recognition .
Cell-Based Detection: LIFR antibodies interacted with cell-derived proteins, enabling functional assays .
Recombinant HRP-Fab conjugates demonstrated enhanced sensitivity in ELISA due to reduced steric hindrance compared to whole antibodies .
| Method | Sensitivity | Equipment Required | Typical Use Case |
|---|---|---|---|
| ELISA | High (chemiluminescent substrates) | Plate reader | Quantitative LIFR analysis |
| Western Blot | Moderate | Imaging system | Protein size confirmation |
The Leukemia Inhibitory Factor Receptor (LIFR) plays a multifaceted role in various biological processes, as evidenced by the following research findings: