trx-2 Antibody
Description
Introduction to TRX-2 Antibody
TRX-2 (Thioredoxin-2) antibodies are specialized immunological tools designed to detect and study the mitochondrial thioredoxin system, a critical regulator of redox homeostasis and apoptosis. These antibodies target TRX-2, a 12–18 kDa protein encoded by the TXN2 gene, which localizes to mitochondria and mitigates reactive oxygen species (ROS) while modulating apoptotic signaling pathways .
3.1. Mechanistic Insights into Mitochondrial Function
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ROS Regulation: TRX-2 antibodies have been pivotal in demonstrating TRX-2’s role in scavenging mitochondrial ROS. Knockout models show elevated ROS levels and cytochrome c release, triggering apoptosis .
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Apoptosis Pathway Modulation: TRX-2 interacts with ASK1 (apoptosis signal-regulating kinase 1), inhibiting its pro-apoptotic activity. Cardiac-specific TRX-2 deletion in mice leads to dilated cardiomyopathy due to ASK1 hyperactivation .
3.2. Disease Models and Therapeutic Targets
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Heart Failure: TRX-2 deficiency in cardiomyocytes correlates with mitochondrial membrane depolarization, reduced ATP production, and progressive heart failure. ASK1 inhibitors reverse these effects in preclinical models .
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Senescence and Aging: Overexpression of TRX-2 in endothelial cells delays oxidative stress-induced senescence, while its SIM (SUMO-interacting motif) mutant loses anti-ROS activity .
4.1. TRX-2 in Cardiac Pathology
4.2. Anti-Senescence and Longevity Studies
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Endothelial Cells: TRX-2 knockdown exacerbates H₂O₂-induced senescence (↑ SA-β-gal activity, p21/p16 expression), while overexpression rescues this phenotype .
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Aging Mice: TRX-2 transgenic mice show reduced mitochondrial ROS and isoprostanes but only marginal lifespan extension (~8–9%) .
5.1. Western Blot Performance
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Observed MW: ~12 kDa (vs. calculated 18 kDa due to mitochondrial processing) .
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Sample Types: Validated in human liver, Raji cells, and mouse tissues .
5.2. Immunohistochemistry (IHC)
Therapeutic Implications
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Basic Research Questions
How do I validate TRX-2 antibody specificity in redox signaling studies?
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Method: Use orthogonal validation techniques:
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Western blot: Confirm absence of bands in TRX-2 knockdown/knockout models .
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Immunoprecipitation: Verify co-precipitation with known TRX-2 interactors (e.g., TXNIP or ASK1) .
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Functional assays: Measure redox activity in samples treated with TRX-2 inhibitors (e.g., PX-12) and compare antibody signal changes .
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What experimental models are optimal for studying TRX-2 in metabolic diseases?
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Preferred systems:
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Primary hepatocytes: For NASH/NAFLD studies, monitor TRX-2 interaction with ASK1 using phospho-specific antibodies (e.g., T838 detection) .
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Endothelial cell lines: Use TNF-α stimulation to assess TRX-2/TXNIP binding dynamics via proximity ligation assays .
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Murine models: Combine TRX-2 KO mice with high-fat diets to analyze metabolic parameter correlations (e.g., insulin resistance) .
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How to design controls for TRX-2 localization studies in complex tissues?
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Critical controls:
Advanced Research Questions
How to resolve contradictions in TRX-2 redox activity measurements across studies?
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Troubleshooting framework:
What strategies improve TRX-2 antibody performance in multiplex assays?
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Optimization steps:
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Epitope mapping: Identify non-overlapping epitopes using HDX-MS to avoid interference with other targets .
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Cross-validation: Pair with redox-sensitive probes (e.g., roGFP2) to correlate antibody signal with actual redox changes .
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Signal amplification: Employ tyramide-based amplification for low-abundance TRX-2 detection in FFPE tissues .
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How to analyze TRX-2/ASK1 interaction dynamics under oxidative stress?
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Protocol:
Methodological Considerations Table
Critical Analysis of Conflicting Data
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Case example: Discrepancies in TRX-2’s role in apoptosis may stem from:
