TGFB2 Antibody
Description
Research Applications of TGFB2 Antibodies
TGFB2 antibodies are validated for diverse experimental workflows:
Immunoassays
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Western Blot (WB): Detects TGF-β2 at ~48–70 kDa in human heart and breast cancer tissues .
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Immunohistochemistry (IHC): Localizes TGF-β2 in formalin-fixed paraffin-embedded (FFPE) samples, such as prostate and colon carcinomas .
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Neutralization: Inhibits TGF-β2-mediated suppression of IL-4-dependent T-cell proliferation (ND₅₀: 0.3–1.5 µg/mL) .
Disease Research
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Cancer: TGF-β2 promotes tumor immunosuppression by converting effector T-cells into regulatory T-cells (T~reg~) . Antibodies like RayBiotech’s 102-10122 are used to study TGF-β2 overexpression in breast and colon cancers .
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Cardiovascular Disorders: Loss-of-function TGFB2 mutations correlate with aortic aneurysms, demonstrated via murine models using antibodies to track TGF-β signaling .
Clinical and Mechanistic Insights
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Cancer Biomarker: TGF-β2 overexpression in tumors correlates with poor prognosis. Antibodies like MAB612 (R&D Systems) identify cytoplasmic TGF-β2 in prostate cancer, linking it to metastasis .
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Fibrosis: TGF-β2 activates hepatic stellate cells (HSCs), driving liver fibrosis. Neutralizing antibodies reduce HSC contractility and collagen deposition .
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Autoimmune Regulation: TGF-β2 upregulates FOXP3 in T~reg~ cells, suppressing anti-tumor immunity. Antibody-blocking studies reveal restored T-cell effector functions .
Technical Considerations
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Cross-Reactivity: Polyclonal antibodies (e.g., AB-12-NA) may cross-react with TGF-β1.2 at high concentrations, while monoclonal variants (e.g., MAB612) show higher specificity .
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Storage: Lyophilized antibodies are stable at -20°C for 12 months; reconstituted aliquots retain activity for 6 months at -70°C .
Product Specs
SASPCCVSQDLEPLTILYYIGKTPKIEQLSNMIVKSCKCS
Q&A
Antibody Selection for TGFB2 Detection
Q: What criteria should guide the selection of TGFB2 antibodies for academic research? A: Selection depends on experimental goals, sample type, and desired specificity. For basic applications (e.g., initial protein detection), polyclonal antibodies like Proteintech’s 19999-1-AP (reactive with human/mouse) or Boster Bio’s A00892 (validated in IHC) are suitable due to broad epitope recognition . For advanced studies (e.g., pathway analysis), recombinant monoclonal antibodies like Proteintech’s 83167-1-PBS (BSA/azide-free) offer batch consistency and conjugation flexibility for multiplex assays . Use host and isotype (e.g., rabbit IgG) to avoid cross-reactivity with secondary antibodies.
Validation of TGFB2 Antibody Specificity
Q: How do I validate TGFB2 antibody specificity in complex biological samples? A: Validate through positive/negative controls and cross-reactivity checks:
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Positive controls: Use cell lines with known TGFB2 expression (e.g., HeLa, A549) .
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Negative controls: Include samples with knocked-out TGFB2 (e.g., CRISPR-edited cells) .
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Cross-reactivity: Test against TGF-β1 or TGF-β3 in WB/IHC to exclude off-target binding .
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Antigen retrieval: Optimize methods (e.g., TE buffer pH 9.0 vs. citrate pH 6.0) for IHC .
Advanced Tip: For signaling pathway studies, confirm colocalization of TGFB2 with SMAD2/3 phosphorylation markers to validate functional relevance .
Optimizing Antibody Dilution for Specific Applications
Q: What dilution ranges are recommended for TGFB2 antibodies in different assays? A: Dilution depends on assay sensitivity and antibody affinity:
Troubleshooting: Weak signals may indicate insufficient primary antibody (try lower dilution) or poor antigen preservation.
Addressing Cross-Reactivity in TGFB2 Assays
Q: How can I mitigate cross-reactivity with other TGF-β isoforms in TGFB2 detection? A: Cross-reactivity is a critical concern due to sequence homology between TGF-β1, TGF-β2, and TGF-β3. Strategies include:
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Epitope specificity: Use antibodies targeting unique regions (e.g., peptide sequences 61-110 for A00892) .
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Isoform-specific controls: Include recombinant TGF-β1/TGF-β3 as negative controls in WB/ELISA .
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Sample pre-treatment: Denature proteins with SDS-PAGE for WB to expose epitopes, reducing non-specific binding .
Advanced Approach: For pathway studies, use phospho-SMAD2/3 antibodies to confirm downstream signaling specificity .
Interpreting Conflicting Data in TGFB2 Studies
Q: How should I resolve discrepancies between TGFB2 antibody results and qPCR data? A: Discrepancies often arise from differences in detection sensitivity or post-translational modifications. Troubleshooting steps:
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Expression vs. secretion: TGFB2 is secreted; antibodies may detect intracellular precursors (latent form) or extracellular mature protein. Use cell lysates vs. conditioned media .
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Protein stability: TGFB2 is prone to degradation; include protease inhibitors (e.g., PMSF) during sample preparation .
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Epitope accessibility: Denatured vs. native epitopes may yield conflicting results (e.g., WB vs. IHC) .
Example: In a study of aortic aneurysms, TGFB2 haploinsufficiency paradoxically elevates TGF-β signaling. Validate findings by comparing TGFB2 protein levels (via antibody) with SMAD2/3 phosphorylation and TGF-β1 ligand expression .
Advanced Applications: Studying TGFB2 in Disease Pathways
Q: How can TGFB2 antibodies be used to study paradoxical TGF-β signaling in vasculopathies? A: TGFB2 haploinsufficiency causes aortic aneurysms despite reduced ligand levels, possibly due to compensatory TGF-β1 upregulation or Ang II crosstalk . Experimental design:
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Mouse models: Use Tgfb2+/− mice to profile SMAD2/3 phosphorylation and TGF-β1 expression in aortic tissue .
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Human samples: Apply IHC with TGFB2 antibodies to thoracic aortic aneurysm specimens to correlate protein loss with fibrotic markers .
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Mechanistic studies: Co-stain for AT1 receptors (Ang II) to explore reciprocal activation pathways .
Troubleshooting Common Issues in TGFB2 IHC
Q: Why do I observe non-specific staining in TGFB2 IHC? A: Non-specific staining often results from:
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Insufficient blocking: Use 5% BSA or non-specific IgG to block Fc receptors .
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Inadequate antigen retrieval: Optimize buffer (TE pH 9.0 vs. citrate pH 6.0) .
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Cross-reactivity: Test with TGF-β1/3 knockouts or peptide-blocking experiments .
Advanced Solution: For complex tissues (e.g., placenta, kidney), use recombinant antibodies (e.g., 83167-1-PBS) with low background .
Comparative Analysis of TGFB2 Antibodies
Q: How do I evaluate the performance of different TGFB2 antibodies? A: Compare via multiplex validation:
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Sensitivity: Detect TGFB2 in serially diluted lysates (e.g., HeLa) .
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Specificity: Use TGF-β isoform-specific controls (e.g., TGF-β1 recombinant) .
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Consistency: Repeat experiments across batches to assess lot-to-lot variability .
Data Example:
| Antibody | WB Sensitivity (ng/mL) | IHC Signal-to-Noise | Cross-Reactivity |
|---|---|---|---|
| 19999-1-AP | 0.5–1.0 | Moderate-High | Human/Mouse |
| 102-10122 | 1.0–2.0 | High | Human |
| 83167-1-PBS | N/A (ELISA-focused) | N/A | Human |
Using TGFB2 Antibodies in Multiplex Assays
Q: How can TGFB2 antibodies be integrated into multiplex platforms? A: Recombinant antibodies (e.g., 83167-1-PBS) enable conjugation with distinct fluorophores or mass tags. Workflow:
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Antibody conjugation: Label with NHS-ester dyes (e.g., DyLight, Alexa Fluor) .
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Panel design: Pair with SMAD2/3, TGF-β1, or Ang II antibodies for pathway profiling .
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Validation: Use cytometric bead arrays or tissue microarrays to confirm specificity .
Ethical and Technical Considerations in TGFB2 Research
Q: What ethical implications should I consider when studying TGFB2 in human tissues? A: Adhere to IRB protocols for human sample use, especially in studies of familial aortic aneurysms . Technical note: Use de-identified specimens and validate findings in orthogonal models (e.g., Tgfb2+/− mice) .
Future Directions for TGFB2 Antibody Development
Q: What innovations are needed to improve TGFB2 antibody performance? A: Prioritize isoform-specific recombinant antibodies and nanobody-based probes for in vivo imaging. Current gaps include antibodies targeting latent vs. active TGFB2 and tools for real-time signaling studies .
Product Science Overview
Transforming Growth Factor-beta 2 (TGF-β2) is a member of the TGF-β family, which plays a crucial role in regulating various cellular processes, including cell growth, differentiation, and immune responses. The TGF-β family is known for its involvement in embryonic development, tissue homeostasis, and the pathogenesis of various diseases, including cancer and fibrosis.
TGF-β2 is a multifunctional cytokine that is involved in the regulation of cell proliferation, differentiation, and apoptosis. It is produced by various cell types, including immune cells, epithelial cells, and fibroblasts. TGF-β2 signals through a receptor complex composed of type I and type II serine/threonine kinase receptors, leading to the activation of intracellular signaling pathways that modulate gene expression.
Polyclonal antibodies are produced by immunizing animals, such as rabbits, with an antigen, in this case, TGF-β2. The immune system of the rabbit generates a diverse population of antibodies that recognize multiple epitopes on the TGF-β2 protein. These antibodies are then collected from the rabbit’s serum and purified for use in various research applications.
The production of polyclonal rabbit anti-human TGF-β2 antibody involves several steps:
- Immunization: Rabbits are immunized with a synthetic peptide corresponding to a specific region of the human TGF-β2 protein. This peptide acts as an immunogen, stimulating the rabbit’s immune system to produce antibodies against TGF-β2.
- Serum Collection: After a series of booster immunizations, blood is collected from the rabbits, and the serum is separated from the blood cells.
- Antibody Purification: The serum contains a mixture of antibodies, including those specific to TGF-β2. These antibodies are purified using techniques such as protein A/G affinity chromatography, which selectively binds to the Fc region of the antibodies, allowing for their isolation from other serum proteins.
The polyclonal rabbit anti-human TGF-β2 antibody is widely used in various research applications, including:
- Western Blotting: To detect and quantify TGF-β2 protein levels in cell and tissue lysates.
- Immunohistochemistry (IHC): To visualize the localization and distribution of TGF-β2 in tissue sections.
- Enzyme-Linked Immunosorbent Assay (ELISA): To measure TGF-β2 concentrations in biological samples, such as serum or cell culture supernatants.
