ITGBL1 Antibody
Description
Definition and Biological Context
ITGBL1 Antibody refers to polyclonal or monoclonal antibodies specifically designed to recognize and bind to the ITGBL1 protein. This protein contains ten integrin-like EGF repeats but lacks transmembrane domains, distinguishing it from classical integrins . ITGBL1 regulates fibrogenesis, epithelial-mesenchymal transition (EMT), and cancer metastasis through pathways like TGF-β/Smad and NF-κB .
Cancer Metastasis Studies
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Colorectal Cancer (CRC): ITGBL1-enriched extracellular vesicles (EVs) promote lung/liver metastasis by activating fibroblasts via NF-κB. Antibodies confirmed elevated ITGBL1 in metastatic CRC tissues (hazard ratio = 2.41 for poor survival) .
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Gastric Cancer (GC): IHC using ITGBL1 antibodies revealed upregulation in GC tissues, correlating with TNM stage and distant metastasis (p < 0.001) .
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Hepatocellular Carcinoma (HCC): ITGBL1 antibodies demonstrated its role in TGF-β/Smad-mediated migration and invasion .
Mechanistic Insights
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Fibroblast Activation: ITGBL1 antibodies identified NF-κB pathway activation in lung fibroblasts treated with CRC-derived EVs .
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EMT Regulation: Western blotting linked ITGBL1 to KRAS/EMT signaling in gastric cancer .
Table 1: Selected Products
| Catalog # | Host | Applications | Reactivity | Supplier |
|---|---|---|---|---|
| NBP1-82473 | Rabbit | IHC, WB | Human, Mouse | Bio-Techne |
| NBP3-02975 | Rabbit | WB | Human, Mouse, Rat | Bio-Techne |
| 30010-1-AP | Rabbit | WB, ELISA | Human | Proteintech |
Table 2: Performance Metrics (Proteintech 30010-1-AP)
| Parameter | Detail |
|---|---|
| Tested Samples | HeLa, SKOV-3 cell lysates |
| Optimal Dilution | WB: 1:500–1:2000 |
| Storage | -20°C in PBS with 50% glycerol |
Limitations and Future Directions
While ITGBL1 antibodies are indispensable for basic research, challenges include cross-reactivity risks (e.g., with integrin β subunits) and limited validation in non-cancer contexts. Future studies should explore ITGBL1’s role in non-malignant fibrotic diseases and optimize antibodies for high-specificity imaging.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Gene References and Functions
- Research has shown that ITGBL1 and its interaction with TGFB1 play crucial roles in regulating fibrogenesis, particularly in the context of hepatitis B-related liver fibrosis. (PMID: 28262670)
- Studies have linked ITGBL1 to cell adhesion. (PMID: 29772438)
- Emerging evidence suggests that ITGBL1 acts as a tumor suppressor in the progression of non-small cell lung cancer. (PMID: 26307393)
- ITGBL1 has potential as a novel biomarker for diagnosing ovarian cancer. (PMID: 27261588)
- Research findings indicate that the transforming growth factor beta (TGFbeta) signaling pathway is a downstream effector of integrin beta-like 1 (ITGBL1) and that the transcription factor Runx2 is an upstream activator of ITGBL1 expression. (PMID: 26060017)
Q&A
How to validate the specificity of ITGBL1 antibodies for different applications?
To ensure antibody specificity, employ peptide competition assays (e.g., pre-incubating antibodies with immunizing peptides to block signal) and positive/negative controls (e.g., using cell lines with known ITGBL1 expression levels) . For Western blotting, confirm molecular weight (~37 kDa) matches ITGBL1’s predicted size, and use reducing/non-reducing conditions to assess protein integrity . In IHC, validate staining patterns in tumor vs. normal tissues, correlating with gene expression data .
What experimental designs optimize ITGBL1 antibody use in studying cancer progression?
For ovarian cancer models, overexpress ITGBL1 in low-expressing cell lines (e.g., SKOV3) and assess adhesion, migration, and chemoresistance via assays like Boyden chamber or cisplatin/paclitaxel IC₅₀ tests . In gastric cancer, combine IHC with EMT markers (e.g., E-cadherin, vimentin) to link ITGBL1 expression to metastasis . For mechanistic studies, pair antibody-based detection with gene knockdown (siRNA/shRNA) to validate functional roles .
How to interpret conflicting ITGBL1 antibody results in IHC?
Discrepancies may arise from antibody epitope differences (e.g., N-terminal vs. central regions) or tissue fixation/processing. Resolve by:
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Comparing staining patterns across antibodies (e.g., cytoplasmic vs. extracellular matrix localization) .
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Correlating IHC scores with mRNA/protein quantification (e.g., qRT-PCR, Western blot) .
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Standardizing protocols (e.g., antigen retrieval conditions, secondary antibody dilutions) .
What advanced methods integrate ITGBL1 antibodies with functional studies?
Proteomics approaches: Use ITGBL1 antibodies in co-IP assays to identify interacting proteins (e.g., integrins, ECM components) influencing cancer cell behavior . Single-cell analysis: Combine flow cytometry with RNA-seq to map ITGBL1 expression to subpopulations with distinct EMT signatures . 3D culture models: Assess ITGBL1’s role in spheroid formation and drug resistance using antibody-based tracking .
How to address cross-reactivity in ITGBL1 antibody applications?
What data analysis strategies resolve ITGBL1’s dual role in cancer?
For prognostic studies, apply Kaplan-Meier survival analysis to correlate ITGBL1 expression levels with OS/DFS, stratifying by clinical subgroups (e.g., metastatic vs. non-metastatic) . In mechanistic studies, employ GSEA to link ITGBL1 to pathways (e.g., KRAS/EMT) and validate with ChIP-seq or luciferase assays .
How to optimize ITGBL1 antibody use in chemoresistance studies?
Experimental workflow:
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Establish dose-response curves for ITGBL1-overexpressing vs. control cells using MTT assays.
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Combine with drug synergy analysis (e.g., cisplatin + paclitaxel) using Combenefit software .
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Validate mechanisms via apoptosis assays (Annexin V/PI) and Western blot for pro-survival proteins (e.g., Bcl-2) .
What computational tools integrate ITGBL1 antibody data with omics platforms?
How to troubleshoot low ITGBL1 antibody signal in Western blot?
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Optimize lysis buffer: Include protease inhibitors (e.g., PMSF) to preserve ITGBL1.
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Normalize loading: Use actin/GAPDH controls and quantify via densitometry.
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Test denaturing conditions: ITGBL1’s EGF-like repeats may require reducing agents (e.g., β-mercaptoethanol) .
What emerging techniques enhance ITGBL1 antibody utility?
Spatial proteomics: Use multiplex IHC with ITGBL1 antibodies to map tumor heterogeneity. CRISPR base editing: Combine with antibody detection to study ITGBL1 gain/loss-of-function. Extracellular vesicle analysis: Isolate EVs and detect ITGBL1 secretion via flow cytometry .
