TNS4 Antibody, HRP conjugated
Description
Definition and Structure
TNS4 Antibody, HRP conjugated, combines a primary antibody targeting the Tensin 4 protein with the HRP enzyme for signal amplification in assays like western blot (WB), immunohistochemistry (IHC), and immunofluorescence (IF). Key structural features include:
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Target: TNS4 (UniProt ID: Q8IZW8), a 77–90 kDa cytoplasmic protein involved in focal adhesion dynamics and tumorigenesis .
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Conjugate: HRP, a 44 kDa glycoprotein that catalyzes chromogenic substrates (e.g., DAB, TMB) for visual detection .
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Host Species: Typically rabbit (polyclonal) or mouse (monoclonal) .
Biological Relevance of TNS4
TNS4 (also called CTEN) regulates cell migration, apoptosis, and epithelial-mesenchymal transition. Its dysregulation is linked to:
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Cancer Progression: Upregulated in colorectal, prostate, and head/neck carcinomas, promoting tumor metastasis .
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Mechanistic Roles:
Conjugation Methodology
HRP conjugation employs site-specific chemistries to preserve antibody affinity. Protocols from Aviva SysBio and Bio-Rad highlight:
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Optimal Antibody Buffer: 10–50 mM amine-free buffer (pH 6.5–8.5) to avoid interference .
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Molar Ratios: Antibody-to-HRP ratios between 1:1 and 1:4 for efficient labeling .
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Stabilizers: Proprietary additives (e.g., LifeXtend™) prevent enzyme degradation in storage .
Conjugation Workflow Table
| Step | Description |
|---|---|
| Antibody Preparation | Dialyze into amine-free buffer (e.g., PBS, HEPES) |
| HRP Mix Activation | Reconstitute lyophilized HRP with modifier reagent |
| Conjugation | Incubate antibody-HRP mixture for 1–2 hours at room temperature |
| Quenching | Add quencher reagent to terminate the reaction |
| Purification | Optional size-exclusion chromatography to remove unbound HRP |
Applications and Validation Data
Validated applications for TNS4-HRP antibodies include:
Table 1: Recommended Dilutions and Applications
| Application | Dilution Range | Observed MW | Validated Cell Lines/Tissues |
|---|---|---|---|
| Western Blot | 1:500–1:2000 | 77–90 kDa | HeLa, HepG2, A549 |
| IHC | 1:20–1:200 | N/A | Human prostate, liver cancer |
| IF/ICC | 1:50–1:500 | N/A | A549 cells |
Key Findings:
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Western Blot: Detects endogenous TNS4 in HepG2 and HeLa lysates .
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IHC: Localizes TNS4 to epithelial cell cytoplasm in prostate tissue .
Research and Clinical Relevance
Recent studies utilizing TNS4-HRP antibodies have revealed:
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Oncogenic Signaling: TNS4 drives colorectal cancer (CRC) progression via β-catenin/c-Myc pathways .
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Therapeutic Target: Suppression of TNS4 enhances sensitivity to cetuximab in KRAS-mutant CRC .
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Prognostic Marker: Elevated TNS4 correlates with poor survival in melanoma and head/neck cancer .
Optimization and Troubleshooting
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- CTEN activated the expression of TGFB1, prompting epithelial-mesenchymal transition in lung adenocarcinoma cancer cells. PMID: 29985912
- Overexpression of the TNS4 gene has been identified as a useful independent predictor of outcomes in stage II/III gastric cancer patients who undergo surgery and receive adjuvant chemotherapy with S-1. PMID: 29102927
- The Cten-Snail signaling pathway contributes to cell motility in colorectal cancer (CRC), mediated by the stabilization of Snail protein. PMID: 28691764
- Research suggests that in colorectal cancer, Cten is upregulated by EGFR and Kras but downregulated by STAT3. PMID: 26852686
- Tensin4 is up-regulated by EGF-induced ERK1/2 activity and promotes cell proliferation and migration in hepatocellular carcinoma. PMID: 26035355
- Cten expression mediates invasion of human lung cancer cells and is upregulated by epidermal growth factor via STAT3. PMID: 25439778
- Findings indicate that induction of Cten protein expression is a relatively early event in melanoma progression, and that Cten has the potential to serve as a prognostic marker for primary melanoma patients. PMID: 24244691
- Up-regulated cten modulates cell migration induced by FGF2. PMID: 23625726
- A significant correlation between MET and TNS4 expression in human colon carcinoma and ovarian carcinoma suggests that TNS4 plays a critical role in MET stability in cancer. PMID: 24814316
- CTEN regulates EGFR protein levels through a posttranslational mechanism. PMID: 23774213
- Phylogenetic analysis, expression patterns, and transcriptional regulation of the human CTEN gene have been studied. PMID: 23500447
- Research has shown an oncogenic role for CTEN in pancreatic cancer through promotion of colony formation and cell motility. PMID: 22750970
- Studies conclude that in the colon and pancreas, Cten is a downstream target of Kras and may be a mechanism through which Kras regulates cell motility. PMID: 21698197
- Cten expression is of prognostic significance in colorectal carcinoma, and a Cten-ILK pathway has been delineated that controls cell motility and possibly promotes metastasis. PMID: 21339732
- CTEN (C-terminal tensin-like) is a novel oncogene that is overexpressed in invasive breast carcinoma. PMID: 20390342
- Increased cten mRNA expression was correlated with tumor progression in lung cancers. PMID: 12711115
- Cten/GAPDH mRNA expression was correlated with evidence of tumor progression in thymoma. PMID: 15001839
- These results provide a novel mechanism whereby the SH2 domain of cten-mediated focal adhesion localization of DLC-1 plays an essential role in its tumor suppression activity. PMID: 17190795
- Expression of CTEN correlates with high tumor grade and metastasis to lymph nodes, suggesting that CTEN may contribute to tumor invasiveness. PMID: 17765673
- TNS4 mRNA expression was significantly higher in gastric cancer. PMID: 18612693
- Cten is a novel nuclear partner of beta-catenin, and has an oncogenic activity in colon cancers. PMID: 19487278
- CTEN plays an important role in paclitaxel sensitivity, and CTEN expression level may be a prognostic predictive factor for prostate cancer patients. PMID: 19725034
