FAF2 Antibody
Description
FAF2 Antibody Overview
FAF2 antibodies are immunoglobulin-based reagents that target the FAF2 protein, which plays roles in apoptosis, lipid metabolism, and organelle homeostasis. These antibodies are available in polyclonal and monoclonal forms, with validated applications across Western blot (WB), immunohistochemistry (IHC), immunoprecipitation (IP), and ELISA .
Apoptosis and Cell Death Regulation
FAF2 antibodies have been critical in elucidating the protein’s role in Fas-mediated apoptosis. Studies show FAF2 interacts with neurofibromin and regulates proteasome-dependent degradation pathways . Knockdown experiments using FAF2-specific shRNA demonstrated reduced hepatic steatosis in ethanol-fed mice, linking FAF2 to lipid metabolism and cell survival .
Peroxisomal Homeostasis
A 2024 study revealed FAF2 as a bifunctional regulator of peroxisomal protein abundance and ether lipid synthesis. FAF2 knockout (KO) cells showed reduced peroxisomal matrix (e.g., AGPS, GNPAT) and membrane proteins (e.g., ABCD3), impairing ether lipid synthesis and sensitizing cells to lipotoxicity .
ER-Associated Degradation (ERAD)
FAF2 antibodies helped identify its role in ERAD, where it facilitates the degradation of misfolded proteins via the ubiquitin-proteasome system. FAF2 interacts with UBQLN2 and YOD1, forming complexes essential for ER stress responses .
Functional Domains and Mechanisms
FAF2 contains two critical domains:
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UBX domain: Mediates interactions with p97/VCP segregase, crucial for peroxisomal regulation .
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UBA domain: Binds ubiquitinated substrates but is dispensable for peroxisomal function .
Mechanistic Insights from KO Studies
| Model System | Key Findings | Citation |
|---|---|---|
| HEK293T cells | FAF2 KO reduced peroxisomal/mitochondrial proteins by 40–60% | BioRxiv (2024) |
| Mouse liver | FAF2 knockdown ↓ PCSK9, ↑ LDLR, ↓ steatosis | PMC (2025) |
Disease Associations
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Cancer: Dysregulated FAF2 is linked to altered apoptosis and proteostasis in tumors .
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Neurodegeneration: FAF2’s role in ERAD implicates it in protein-misfolding diseases .
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Alcoholic Liver Disease (ALD): FAF2 silencing reduced hepatic lipid accumulation via ATGL-mediated lipolysis and SREBP1/PCSK9 modulation .
Therapeutic Targeting
FAF2 antibodies are used to validate its potential as a drug target. For example:
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Liver Disease: AAV8-mediated FAF2 knockdown in mice reduced ethanol-induced steatosis by 50% .
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Lipotoxicity: FAF2 inhibitors could mitigate peroxisome dysfunction in metabolic disorders .
Challenges and Future Directions
Product Specs
Target Background
Q&A
Basic Research Questions
What criteria should guide FAF2 antibody selection for Western blotting?
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Prioritize antibodies with validated molecular weight specificity (e.g., 53 kDa for human FAF2 ) and cross-reactivity across species (human, mouse, rat).
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Compare validation data across vendors (Table 1):
Methodological Guidance:
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Include knockout controls (e.g., FAF2-/- cells ) to confirm specificity.
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Optimize dilution ranges (e.g., 1:500–1:50,000 for WB ) using titration protocols.
How can researchers validate FAF2 antibody specificity in immunohistochemistry (IHC)?
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Use antigen retrieval buffers (e.g., TE buffer pH 9.0 or citrate buffer pH 6.0 ) to enhance epitope accessibility.
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Validate with tissue-specific negative controls (e.g., FAF2-/- mouse brain tissue ) and correlate with mRNA/protein expression databases.
Advanced Research Questions
How to resolve discrepancies in FAF2 subcellular localization across studies?
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Context-dependent localization: FAF2 exhibits dual roles in peroxisomal homeostasis and ER-associated degradation . Use subcellular fractionation paired with organelle-specific markers (e.g., PMP70 for peroxisomes ).
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Technical considerations:
What experimental strategies elucidate FAF2’s role in lipid metabolism?
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CRISPR/Cas9 screens: Genome-wide sgRNA libraries (e.g., Brunello library) can identify FAF2-dependent pathways under lipotoxic stress .
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Functional assays:
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Transcriptomic analysis: Perform GSEA on RNA-seq data from FAF2-knockdown models to identify enriched pathways (e.g., AMPK signaling, fatty acid degradation ).
How to address inconsistent FAF2 detection in Western blots?
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Post-translational modifications: FAF2 undergoes phosphorylation under stress; use phosphatase inhibitors in lysis buffers.
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Sample preparation: Avoid overloading lysates from lipid-rich tissues (e.g., liver), which may mask FAF2 bands .
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Antibody lot validation: Compare performance across lots using reference lysates (e.g., HEK-293 cells ).
What methodologies confirm FAF2’s role in apoptosis regulation?
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Co-immunoprecipitation (Co-IP): Use antibodies validated for IP (e.g., 16251-1-AP ) to isolate FAF2 complexes with apoptosis regulators (e.g., Fas-associated factors).
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Functional rescue experiments: Overexpress FAF2 in knockout models (e.g., Flag-FAF2 plasmids ) and assess apoptosis via propidium iodide/annexin V staining.
Data Contradiction Analysis
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Discrepant molecular weights: Some studies report 51 kDa vs. 53 kDa . This may arise from splice variants or degradation; always include recombinant FAF2 controls.
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Opposing roles in steatosis: Hepatic FAF2 knockdown reduces lipid accumulation , while renal models show peroxisomal dysregulation . Context-specific analyses (e.g., tissue-specific Cre models) are critical.
