TARDBP Antibody
Description
Introduction to TARDBP Antibody
TARDBP antibodies are immunological tools designed to detect TAR DNA-binding protein 43 (TDP-43), a multifunctional RNA/DNA-binding protein encoded by the TARDBP gene. These antibodies are critical for studying TDP-43's roles in transcriptional regulation, RNA splicing, and disease pathogenesis, particularly in neurodegenerative disorders like amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD), as well as cancer .
Association with Neurodegenerative Diseases
TDP-43 pathology is hallmark of ALS and FTLD-U, where cytoplasmic inclusions of phosphorylated TDP-43 disrupt neuronal function . Key findings:
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ALS Patients: Serum anti-TDP-43 autoantibodies are elevated 85% compared to controls, independent of TARDBP mutations .
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Diagnostic Utility: Antibodies like Proteintech’s 10782-2-AP (targeting N-terminal TDP-43) and 12892-1-AP (C-terminal) detect pathological forms in brain tissues .
Role in Cancer Biology
TDP-43 exhibits dual oncogenic and tumor-suppressive roles depending on cancer type:
Key Research Findings and Clinical Implications
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Pan-Cancer Analysis: TDP-43 is universally upregulated in tumors (e.g., THYM, TGCT) and correlates with tumor mutational burden (TMB) and immune evasion .
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Immune Microenvironment: Negative correlation with StromalScore/ImmuneScore in 17 cancers, suggesting immunosuppressive effects .
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Therapeutic Target: In HCC, TDP-43 promotes proliferation and invasion, validated via CCK8 and Transwell assays .
Validation and Performance of TARDBP Antibodies
A 2023 study evaluated 18 commercial TDP-43 antibodies for specificity and reproducibility :
| Application | Top-Performing Antibodies | Clone/Product | Validation Outcome |
|---|---|---|---|
| Western blot | Proteintech 10782-2-AP, R&D Systems MAB7778 | Rabbit polyclonal, Mouse monoclonal | Specific bands at 43–45 kDa in KO-validated cell lines |
| Immunofluorescence | R&D Systems MAB77781 | Mouse monoclonal | Nuclear localization in splenocytes and brain tissues |
| Immunoprecipitation | Abnova H00023435-M01 | Clone 2E2-D3 | Efficient pull-down of TDP-43 from HAP1 lysates |
Product Specs
Q&A
How do I select a TARDBP antibody for Western blotting in murine models?
Optimal antibody selection requires evaluating species reactivity, epitope specificity, and validation data. For murine systems, the TARDBP Antibody (E-10) (sc-376311) recognizes mouse, rat, and human isoforms due to conserved epitopes within the N-terminal Met1-Thr103 region . Prioritize antibodies validated in knockout (KO) models, as demonstrated in HAP1 cell studies where WT vs. TARDBP KO lysates confirmed specificity . Key parameters:
What controls are essential for TARDBP immunohistochemistry (IHC)?
Robust IHC requires:
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Negative Controls: TARDBP KO tissue sections or siRNA-treated cells to confirm signal absence .
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Competition Assays: Pre-incubation with recombinant TDP-43 protein (e.g., Met1-Thr103 fragment) to block specific binding .
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Multi-Target Validation: Co-staining with RNA-binding proteins like FUS or HNRNPA1 to exclude nonspecific nuclear aggregation .
Why does TARDBP appear as multiple bands in Western blots?
TARDBP’s post-translational modifications (phosphorylation, ubiquitination) and alternative splicing generate isoforms between 35-45 kDa. To resolve this:
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Use urea-SDS-PAGE (4–12% gradient gels) to separate phosphorylated (p-TDP-43) and truncated forms .
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Include phosphatase inhibitors (e.g., PhosSTOP) during lysis to preserve modification states .
How can I resolve contradictions in TARDBP subcellular localization data?
Discrepancies (nuclear vs. cytoplasmic staining) often arise from:
In cancer studies, cytoplasmic TARDBP correlates with elevated Tumor Mutational Burden (TMB), necessitating dual IF/WB validation .
What genetic models are optimal for studying TARDBP loss-of-function?
CRISPR-engineered HAP1 cells provide a standardized system:
| Cell Line | Genotype | Application | Source |
|---|---|---|---|
| HAP1 WT | TARDBP +/+ | Baseline expression controls | Horizon Discovery |
| HAP1 KO | TARDBP −/− | Specificity validation | CVCL_TR64 |
For in vivo models, Tardbp heterozygous mice exhibit age-dependent motor deficits, mimicking ALS progression .
How does TARDBP influence immune checkpoint regulation in oncology?
In hepatocellular carcinoma (HCC), TARDBP upregulation inversely correlates with PD-L1 and CTLA-4 expression (Spearman’s ρ = −0.32, p < 0.01), suggesting immune evasion suppression. Methodological insights:
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ESTIMATE Algorithm: Compute ImmuneScores (Stromal/Immune cell ratios) using TCGA data .
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Multiplex IF: Pair TARDBP antibodies with CD8+/FoxP3+ markers to map tumor-infiltrating lymphocytes .
Can TARDBP antibodies be used in multiplex RNA-protein co-detection assays?
Yes, but with protocol adjustments:
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Sequential Staining: Perform RNA FISH (e.g., CFTR exon 9 probes) before TARDBP IHC to prevent epitope masking .
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Crosslinking Reagents: Use DSG (disuccinimidyl glutarate) to stabilize RNA-protein interactions during IF .
What computational tools enhance TARDBP interaction network analysis?
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STRING-DB: Predicts TARDBP-binding partners (e.g., HNRNPA1, FUS) with confidence scores >0.7 .
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Cytoscape Plugins: iRegulon identifies TARDBP-regulated genes using ChIP-seq data from ENCODE .
How do phosphorylation states affect TARDBP antibody performance?
Phospho-specific antibodies (e.g., pS409/410) require:
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Phosphatase Treatment: Pre-lysis treatment with λ-PPase to distinguish phosphorylated vs. total TDP-43 .
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2D Gel Electrophoresis: Separate isoforms by charge (pI 5.8 vs. 6.2) before Western blotting .
What are the pitfalls in correlating TARDBP expression with clinical outcomes?
Common biases:
Product Science Overview
TAR DNA-binding protein 43 (TDP-43) is a crucial RNA-binding protein that plays a significant role in RNA processing and stability. It was initially identified as a transcriptional repressor binding to the TAR DNA element of the HIV-1 virus . TDP-43 has since been shown to bind both RNA and DNA, and it has multiple functions in transcriptional repression, translational regulation, and pre-mRNA splicing .
In 2006, TDP-43 was identified as a major component of ubiquitinated inclusions in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) . It was found to be mislocalized from the nucleus to the cytoplasm in neurons and glia in affected central nervous system regions . Soon after, TDP-43 mutations were discovered in families with inherited ALS and FTD, suggesting a primary role for TDP-43 disruption in disease pathogenesis .
The nuclear clearance and cytoplasmic mislocalization of TDP-43 are pathological hallmarks of ALS, FTD, and related neurodegenerative disorders collectively termed "TDP-43 proteinopathies" . TDP-43 mislocalization causes neurodegeneration through both loss and gain of function mechanisms. Loss of TDP-43 nuclear RNA processing function destabilizes the transcriptome by multiple mechanisms, including disruption of pre-mRNA splicing, failure of repression of cryptic exons, and retrotransposon activation . The accumulation of cytoplasmic TDP-43, which is prone to aberrant liquid–liquid phase separation and aggregation, traps TDP-43 in the cytoplasm and disrupts a host of downstream processes, including the trafficking of RNA granules, local translation within axons, and mitochondrial function .
The mouse anti-human TDP-43 antibody is a monoclonal antibody that specifically detects human TDP-43 in various applications, including Western blot, immunohistochemistry, immunoprecipitation, and immunocytochemistry . This antibody is purified from hybridoma culture supernatant using protein A or G and is available in lyophilized form . It is used in research to study the expression and localization of TDP-43 in human, mouse, and rat tissues .
The mouse anti-human TDP-43 antibody has been widely used in research to investigate the role of TDP-43 in neurodegenerative diseases. It helps in understanding the mechanisms associated with TDP-43 neurotoxicity in ALS and FTD . Researchers use this antibody to study the mislocalization and aggregation of TDP-43 in affected neurons and glia, providing insights into the disease mechanisms and potential therapeutic targets .
