Rbfox2 Antibody
Description
Research Applications of Rbfox2 Antibodies
Rbfox2 antibodies are widely used in experimental workflows. Key applications include:
Antibodies such as HPA006240 and CSB-PA019439LA01HU are commercially available, with conjugates (HRP, FITC, biotin) optimized for ELISA and multiplex assays .
Acute Myeloid Leukemia (AML)
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Oncogenic Function: RBFOX2 is overexpressed in AML subtypes and associated with poor survival. Depletion reduces leukemic stem cell (LSC) self-renewal, promotes differentiation, and prolongs survival in PDX models .
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Mechanism: RBFOX2 recruits RBM15 to enhance m6A methylation on caRNAs, repressing tumor suppressors like TGFB1. RBFOX2 knockdown upregulates TGF-β signaling, inducing myeloid differentiation .
Solid Tumors
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Pancreatic Ductal Adenocarcinoma (PDA): RBFOX2 acts as a metastasis suppressor by regulating alternative splicing of Rho GTPase pathway genes (e.g., RHOA, CDC42). Overexpression reduces colony formation and lung metastases in vivo .
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Hepatocellular Carcinoma (HCC): Low RBFOX2 expression correlates with increased proliferation and metastasis. Knockdown enhances EMT and invasiveness .
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Immune Microregulation: RBFOX2 expression inversely correlates with CD8+ T-cell infiltration and positively associates with immunosuppressive cells (CAFs, MDSCs), suggesting a role in immune evasion .
Key Research Findings
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Leukemia Maintenance: RBFOX2 depletion in AML reduces LSC frequency (1/169.41 vs. 1/6.28 in controls, P < 0.001) and inhibits leukemogenesis in PDX models .
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Splicing Regulation: In PDA, RBFOX2 modulates >87 splicing targets, including MPRIP and TEAD1, to suppress metastasis .
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Immunomodulatory Effects: Pan-cancer analysis shows RBFOX2 inversely correlates with immune checkpoint genes (e.g., PD-L1, CTLA-4), suggesting therapeutic implications .
Future Directions
RBFOX2 antibodies will remain vital for studying RNA-protein interactions and therapeutic targeting. Ongoing research focuses on:
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Gene References and Functions
- Recent research has revealed a distinct alternative splicing program activated by Rbfox2 in endothelial cells during platelet and macrophage recruitment. This program plays a crucial role in transcriptional responses during flow-driven vascular inflammation. PMID: 29293084
- A study demonstrated that Rbfox2 represses the inclusion of exon 18a during the pre-mRNA splicing of CaV2.2, a voltage-gated calcium channel. This regulation restricts the magnitude of CaV2.2 currents during early development in specific neuronal populations. PMID: 29067356
- Dysregulation of Rbfox2, particularly by dominant-negative Rbfox2, has been identified as an early pathogenic event in diabetic hearts. PMID: 27239029
- Rbfox2 interacts with chromatin in a manner dependent on nascent RNA. Inactivation of Rbfox2 disrupts the targeting of Polycomb Repressive Complex 2 (PRC2) to a majority of bivalent gene promoters, leading to transcriptional derepression. PMID: 27211866
- A classic nuclear localization signal at the N terminus of Rbfox2 has been identified, regulating the subcellular localization of Rbfox2 isoforms during the differentiation of mouse epithelial and embryonic stem cell lines. PMID: 27859055
- Gain- and loss-of-function experiments have demonstrated that Rbfox1 and Rbfox2 collaborate to promote Mef2D splicing, subsequently contributing to myogenesis (muscle cell development). PMID: 25609712
- Research indicates that the conserved Rbfox2 RNA binding protein regulates approximately 30% of the splicing transitions observed during myogenesis. Importantly, Rbfox2 is essential for the specific step of myoblast fusion, a crucial process in muscle development. PMID: 25087874
- Studies have unveiled an unexpectedly broad and multifaceted regulatory network controlled by Rbfox2. This network provides an explanation for how autoregulatory splicing networks are precisely tuned. PMID: 24637117
- During epithelial-mesenchymal transition (EMT), a process involved in tissue remodeling, Rbfox2-regulated splicing shifts from epithelial-specific to mesenchymal-specific events. PMID: 23435423
- Rbfox2 protein plays a critical role in controlling a post-transcriptional program essential for proper brain development. PMID: 22357600
- Research has elucidated a novel mechanism by which Rbfox3 negatively regulates Rbfox2. PMID: 21747913
- A study demonstrated that alternative neuronal isoforms of the splicing factors nPTB and Fox-1/Fox-2 are also produced in lenses. PMID: 21714144
- Fox-1 and Fox-2 isoforms specifically activate the splicing of neuronally regulated exons. This activation requires the presence of UGCAUG enhancer elements within the RNA sequence. PMID: 16260614
Q&A
Basic Research Questions
How to validate Rbfox2 antibody specificity in Western blotting for embryonic stem cell studies?
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Perform knockout controls using Rbfox2 conditional knockout mouse models (e.g., Nkx2.5 Cre system) . Compare lysates from wild-type and mutant tissues (e.g., heart or ESC-derived cells) to confirm absence of target bands in mutants.
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Use siRNA-mediated knockdown in cell lines (e.g., HEK293) paired with FLAG-tagged RBFOX2 overexpression to distinguish endogenous vs. exogenous protein .
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Validate cross-reactivity with other Rbfox paralogs (e.g., Rbfox1/3) using isoform-specific peptide blocking assays.
What experimental parameters optimize Rbfox2 antibody performance in immunofluorescence?
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Fixation: 4% PFA for 15 min preserves epitopes for nuclear/cytoplasmic localization studies .
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Permeabilization: 0.3% Triton X-100 for 10 min balances membrane integrity and antibody penetration.
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Antigen retrieval: Citrate buffer (pH 6.0) at 95°C for 20 min improves signal for formalin-fixed paraffin-embedded samples .
How to address Rbfox2 isoform detection challenges due to alternative splicing-coupled NMD?
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Combine RNA-seq data with antibody validation to correlate protein expression with splice variants .
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Use cycloheximide (50 µg/ml, 4 hr pretreatment) to inhibit NMD and amplify truncated isoforms .
Advanced Research Questions
Resolving contradictory Rbfox2 localization data across studies: nuclear vs. cytoplasmic
Designing multiplexed assays to study Rbfox2-RNA interaction networks
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Integrate iCLIP-seq (UV crosslinking) with co-immunoprecipitation (IP) using RNAse inhibitor-treated lysates .
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Protocol:
Validating Rbfox2 antibody specificity in disease models with altered splicing landscapes
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Heart failure models: Compare Rbfox2 expression in pressure-overload hypertrophy vs. controls using quantitative IF and RNA-FISH .
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Data normalization: Use housekeeping genes with stable splicing (e.g., Gapdh constitutive exons) to control for global splicing changes .
Methodological Considerations Table
Key Validation Data from Literature
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Antibody Clone Cross-reactivity
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Dynamic Range in Disease States
