sre-8 Antibody
Description
Definition and Biological Role of Sur-8 Antibody
Sur-8 Antibody (D-8) is a mouse-derived monoclonal IgG1κ antibody targeting the Sur-8 protein (SHOC2) in humans, mice, and rats. It detects Sur-8 via multiple methods, including Western blotting (WB), immunoprecipitation (IP), immunofluorescence (IF), and ELISA .
Sur-8 is a 582-amino-acid cytoplasmic protein containing 20 leucine-rich repeats (LRRs) that facilitate protein-protein interactions. It dynamically translocates to the nucleus upon growth factor stimulation, modulating Ras-MAPK signaling . Dysregulation of Sur-8 due to aberrant N-myristoylation is linked to Noonan-like syndrome with loose anagen hair, characterized by developmental delays and hair abnormalities .
Antibody Structure
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Epitope: Binds to Sur-8’s LRR domain, critical for its scaffolding role in Ras-MAPK signaling .
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Conjugates: Available in agarose, HRP, PE, FITC, and Alexa Fluor® formats for multiplex assays .
Target Protein Features
| Feature | Description |
|---|---|
| Molecular Weight | ~82 kDa (predicted) |
| Domains | 20 LRRs, nuclear localization signal |
| Cellular Localization | Cytoplasm (basal), nucleus (post-stimulation) |
| Pathway | Ras-MAPK signaling (cell proliferation, differentiation) |
Experimental Utility
Sur-8 Antibody (D-8) has been validated in:
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Western Blotting: Detects endogenous Sur-8 in HEK293 and HeLa cell lysates .
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Immunoprecipitation: Isolates Sur-8 complexes for pathway analysis .
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Immunofluorescence: Visualizes Sur-8 translocation in response to EGF stimulation .
Performance in RAS Pathway Studies
A large-scale antibody validation study for RAS pathway components reported:
| Application | Tested Antibodies | Positive Results | Validation Matrix |
|---|---|---|---|
| Western Blotting | 119 | 63 | Recombinant protein |
| Western Blotting | 63 | 41 | Cell lines |
| Immunoprecipitation | 61 | 56 | Recombinant protein |
This underscores the rigorous validation required for antibodies targeting signaling proteins like Sur-8 .
Role in Disease
Sur-8 mutations disrupt Ras-MAPK signaling, leading to:
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Noonan-like syndrome: Mutations (e.g., SHOC2-S2G) cause ectopic localization and hyperactivity .
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Cancer: Overexpression correlates with tumor progression in glioma and melanoma models.
In Vivo and In Vitro Findings
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Knockdown Studies: Sur-8 depletion inhibits Ras-driven oncogenesis in cellular assays .
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Therapeutic Targeting: Sur-8 is explored as a biomarker for MAPK inhibitor resistance .
Limitations and Future Directions
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
KEGG: cel:CELE_F40G12.1
UniGene: Cel.3639
Q&A
FAQs for Sur-8 (SHOC2) Antibody Research
Advanced Research Questions
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How should experimental design account for Sur-8’s role in neutrophil recruitment and fibrosis?
Sur-8 interacts with IL-8 signaling pathways, which influence inflammation and fibrosis (e.g., endometriosis models ). Considerations include:-
In vivo models: Use humanized mice reconstituted with myeloid cells to study Sur-8’s impact on neutrophil chemotaxis .
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Combination therapies: Pair Sur-8 inhibitors with PD-1 blockers to assess synergistic effects on tumor microenvironments .
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Dosage optimization: For monoclonal antibodies, refer to concentration guidelines (e.g., 1–5 µg/ml for IHC ).
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How to resolve contradictions in Sur-8 antibody performance across studies?
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What advanced techniques leverage Sur-8 antibodies for structural biology?
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Crystallography: Co-crystallize Sur-8 Fab fragments with binding partners (e.g., Ras) to map interaction interfaces .
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Neutralization assays: Test Sur-8’s role in blocking Ras-MAPK signaling using competitive ELISAs .
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Single-cell sequencing: Correlate Sur-8 expression with Ras pathway activation in tumor subpopulations .
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Methodological Best Practices
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Antibody storage: Aliquot Sur-8 antibodies (mouse monoclonal IgG1κ) at -80°C to prevent freeze-thaw degradation .
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Concentration guidelines:
Application Recommended Concentration Western blot 1:500–1:1000 dilution Immunofluorescence 1:50–1:200 dilution ELISA 0.5–2 µg/ml -
Data interpretation: Use computational tools (e.g., STRING database) to contextualize Sur-8 within Ras-MAPK interaction networks .
