FXR2 Antibody
Description
Overview of FXR2 Protein
FXR2 (UniProt ID: P51116) is a member of the Fragile X protein family, sharing structural homology with FMRP and FXR1. It regulates mRNA stability and translation through interactions with polyribosomes and RNA molecules . Key features include:
-
Functions:
Development of FXR2 Antibodies
Commercial FXR2 antibodies are primarily rabbit-derived polyclonal or monoclonal reagents. Notable examples include:
| Product | Host | Applications | Reactivity | Catalog # |
|---|---|---|---|---|
| Rabbit Anti-FXR2 | Rabbit | WB, IHC, IF/ICC | Human, Mouse | DF3195 |
| FXR2 Polyclonal Antibody | Rabbit | WB, IP, RIP, ELISA | Human, Mouse | 12552-1-AP |
Implications:
-
FXR2 may serve as an indirect biomarker for HER2-negative BC .
-
High FXR2 correlates with hormone receptor negativity, suggesting a role in BC subtypes .
Technical Considerations
-
Western Blot: Detects bands at ~75–115 kDa due to isoform diversity .
-
Immunohistochemistry: Optimized protocols recommend antigen retrieval with TE buffer (pH 9.0) .
-
Controls: Use HEK-293T or HeLa cell lysates for validation .
Research Tools and Reagents
Recombinant FXR2 proteins are available for antibody validation :
| Product | Source | Tag | Species |
|---|---|---|---|
| FXR2-13053H | E. coli | His-tagged | Human |
| FXR2-1593R | HEK293 | Fc/His/Avi | Rhesus macaque |
Future Directions
Product Specs
Target Background
Customer Reviews
Applications : Co-immunoprecipitation assay and Immunofluorescence
Sample type: cell
Review: Co-immunoprecipitation assay indicated a direct interaction of FXR and SIRT6; SIRT6 overexpression decreases the acetylation of FXR.
Q&A
How can researchers validate the specificity of FXR2 antibodies in rodent models?
Validation requires a multi-step approach:
-
Knockout (KO) controls: Use Fxr2 KO mouse brain lysates (e.g., from studies in ) to confirm absence of signal in WB/IHC.
-
Cross-reactivity testing: Compare antibody performance against FMR1 and FXR1 using recombinant proteins (e.g., FXR2 Antibody 1G2 shows no cross-reactivity with FMR1 in WB ).
-
Orthogonal methods: Combine IP-MS with RNA-binding assays to confirm target specificity (e.g., RIP-seq in identified PSD-95 as a validated FXR2 target).
Table 1: Common Validation Strategies for FXR2 Antibodies
What experimental conditions optimize FXR2 detection in Western blotting?
-
Sample preparation: Use RIPA buffer with RNase inhibitors to preserve RNA-protein complexes ( ).
-
Gel percentage: 8–12% SDS-PAGE gels resolve FXR2’s observed molecular weight (~95 kDa, higher than calculated 74 kDa due to post-translational modifications ).
-
Antibody dilution: 1:1,000–1:4,000 for polyclonal antibodies (e.g., ab168852 ); 2–4 μg/mL for monoclonal clones (e.g., A42 ).
Critical Parameters:
-
Blocking agents: 5% BSA reduces non-specific binding in neuronal lysates ( ).
-
Antigen retrieval: TE buffer (pH 9.0) enhances epitope exposure in FFPE tissues ( ).
How can FXR2 be distinguished from FMR1/FXR1 in co-immunoprecipitation studies?
-
Antibody selection: Use isoform-specific clones (e.g., 1G2 for FXR2 vs. 7G1 for FMR1 ).
-
Genetic controls: Compare IP results in Fmr1 KO vs. Fxr2 KO models ( ).
-
Bioinformatic analysis: Overlap RNA targets identified via RIP-seq with known FMR1/FXR1 interactomes ( ).
What methodologies identify FXR2’s role in phase-separated membraneless compartments?
-
Liquid-liquid phase separation (LLPS) assays: Treat cells with 1,6-hexanediol to disrupt phase-separated structures and monitor FXR2 redistribution ( ).
-
Single-molecule FISH: Combine with IF to colocalize FXR2 with AU-rich element (ARE)-containing mRNAs (e.g., NOG ).
-
Live-cell imaging: Track GFP-tagged FXR2 dynamics under stress conditions (e.g., Zika virus infection ).
Key Findings:
-
FXR2 phase separation is RNA-dependent and regulates mRNA storage/translation in hippocampal neurons ( ).
-
SARS-CoV-2 hijacks FXR2-mediated LLPS to cluster replication organelles ( ).
How does FXR2 modulate ERK1/2 signaling in kainate-induced seizure models?
-
Kinase activity profiling: Compare phospho-ERK levels in WT vs. Fxr2 KO mice post-kainic acid (KA) injection ( ).
-
Electrophysiology: Measure hippocampal burst activity using multi-electrode arrays (reduced in Fxr2 KO ).
-
Transcriptomic analysis: Identify FXR2-bound mRNAs encoding glutamatergic receptors (e.g., GRIA2, GRIN2A) via RIP-seq ( ).
Mechanistic Insights:
-
FXR2 stabilizes mRNAs for postsynaptic density proteins, sustaining ERK phosphorylation required for seizure propagation ( ).
What techniques confirm FXR2’s interaction with Zika virus (ZIKV) sfRNA?
-
RNA-protein pull-down: Use biotinylated sfRNA probes in ZIKV-infected HeLa cells ( ).
-
Flow cytometry: Quantify FXR2 expression in infected vs. uninfected cells (2-fold increase in WT ZIKV ).
-
Functional assays: Compare viral replication efficiency in Fxr2 KO cells (e.g., plaque assays ).
Pathological Relevance:
-
ZIKV sfRNA sequesters FXR2, dysregulating translation of synaptic proteins (e.g., PNPLA6) and exacerbating neurodevelopmental defects ( ).
