TNN Antibody, HRP conjugated
Description
Introduction to TNN Antibody, HRP Conjugated
The TNN Antibody, HRP conjugated, refers to a horseradish peroxidase (HRP)-labeled antibody targeting the Tn antigen, a carbohydrate epitope (GalNAc-α-O-Ser/Thr) commonly expressed on tumor cells. This conjugate is widely used in immunoassays, such as ELISA and immunohistochemistry (IHC), for detecting Tn antigen in cancer diagnostics and research. The HRP enzyme enables chromogenic or luminescent detection via substrates like diaminobenzidine (DAB) or TMB, producing measurable signals proportional to antigen presence .
Conjugation Methods and Stability
2.1. Classical Conjugation
HRP is typically conjugated to antibodies via periodate-mediated oxidation of the enzyme’s carbohydrate moieties, creating aldehyde groups that react with lysine residues on the antibody. This method often requires optimized buffer conditions (e.g., avoiding reducing agents like sodium azide) to prevent interference with conjugation .
2.2. Enhanced Conjugation via Lyophilization
A modified protocol involves lyophilizing activated HRP after periodate treatment, reducing reaction volume while maintaining reactant concentrations. This step increases the number of HRP molecules bound per antibody, enhancing sensitivity. For example, lyophilized conjugates achieved ELISA detection at 1:5000 dilution compared to 1:25 for classical methods (p < 0.001) .
| Parameter | Classical Method | Lyophilized Method |
|---|---|---|
| Antibody Dilution | 1:25 | 1:5000 |
| Sensitivity | Moderate | High |
| Storage Stability | Short-term | Long-term (4°C) |
Applications in Cancer Research
3.1. ELISA and IHC Detection
The TNN Antibody, HRP conjugated, is used to detect Tn antigen in tumor tissues. In breast cancer studies, conjugates showed specific binding to MCF7 cells, with in vivo imaging confirming localization to primary tumors and metastases .
3.2. Therapeutic Potential
Anti-Tn antibodies inhibit tumor cell adhesion to lymphatic endothelial cells (LECs), suggesting a role in preventing lymphatic metastasis. Conjugation with HRP facilitates tracking of antibody distribution in preclinical models .
Research Findings and Data
4.1. ELISA Performance
A study comparing conjugation methods demonstrated superior sensitivity for lyophilized conjugates:
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Signal-to-Noise Ratio: 3.5-fold higher for lyophilized conjugates at 1:5000 dilution .
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Antigen Binding: SDS-PAGE confirmed efficient conjugation, with non-reducing gels showing no migration of conjugated products .
4.2. In Vivo Imaging
HRP-conjugated anti-Tn antibodies, combined with QDot labeling, enabled real-time tumor visualization in mice. Tumor-to-background ratios exceeded 5:1, validating targeting efficiency .
| ELISA Parameter | Classical Conjugate | Lyophilized Conjugate |
|---|---|---|
| OD450 (1:25) | 0.8 ± 0.1 | 1.2 ± 0.2 |
| OD450 (1:5000) | N/A | 0.95 ± 0.15 |
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Tenascin-W is an extracellular matrix protein that functions as a ligand for integrins α8β1, αVβ1, and α4β1. It plays a role in neurite outgrowth and cell migration within hippocampal explants. In endochondral bone formation, it inhibits the proliferation and differentiation of proteoblasts via modulation of canonical Wnt signaling. Furthermore, in tumor environments, Tenascin-W stimulates angiogenesis by promoting the elongation, migration, and sprouting of endothelial cells. Its expression is prevalent in many mammary tumors, potentially contributing to tumorigenesis by facilitating the migratory behavior of breast cancer cells.
Supporting Research:
- A study identified potential disease-causing mutations in the TTN gene in 21 patients, clarifying the genetic basis of their condition in at least part. PMID: 26627873
- Research indicates that Tenascin-W supports breast cancer metastasis to bone by promoting cancer cell migration and proliferation. PMID: 25868708
- Tenascin-W expression in mammary tumors is regulated by p38MAPK and JNK signaling pathways. PMID: 15592496
- Studies suggest that Tenascin-W expression in activated tumor stroma promotes tumorigenesis by supporting breast cancer cell migration. PMID: 17909022
- Elevated Tenascin-W levels are correlated with the presence of colorectal and breast cancer. PMID: 18306355
