MET4 Antibody
Description
Introduction to MET4 Antibody
The MET4 Antibody (clone 8G6) is a mouse-derived monoclonal antibody designed to recognize an extracellular epitope of human c-MET . It was specifically engineered for compatibility with formalin-fixed, paraffin-embedded (FFPE) tissues, enabling standardized immunohistochemical (IHC) evaluation in clinical diagnostics . c-MET overexpression correlates with poor prognosis in cancers, making MET4 a vital tool for patient stratification .
Antibody Generation
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Derived from hybridoma technology using recombinant MET proteins for immunization .
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Selected for its distinct staining pattern in normal prostate tissues, showing specificity for luminal cell membranes .
Technical Performance
MET4 demonstrated 100% concordance with immunoblotting in FFPE cell pellets, confirming accuracy .
Ovarian Cancer and Glioma
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Ovarian Cancer: 25% exhibited strong MET4 staining across histological subtypes .
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Glioma: 63% positivity, suggesting utility in neuro-oncology .
Companion Diagnostic Utility
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Licensed to Dako for global distribution as a pharmDx™ assay .
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Validated for fluorescence-activated cell sorting (FACS) in detecting surface c-MET on SK-LMS-1/HGF and MNNG-HOS cell lines .
Comparative Advantages Over Existing Antibodies
| Feature | MET4 | SP44/D1C2 (Intracellular) | C-28 Polyclonal |
|---|---|---|---|
| Epitope Location | Extracellular domain | Cytoplasmic domain | C-terminus |
| FFPE Compatibility | High (optimized protocols) | Moderate | Variable |
| Correlation with p-MET | Strong (P < 0.001) | Weak | Not assessed |
| Stromal Background | Low | High | High |
MET4’s extracellular targeting enables precise detection of membranous c-MET, avoiding cytoplasmic false positives .
Future Directions
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
MET4 Antibody Research FAQs
Advanced Research Questions
How to resolve MET4 binding discrepancies under varying sulfur conditions?
| Condition | Binding Pattern | Resolution Method |
|---|---|---|
| Sulfur-replete | Cbf1-dependent promoters only | ChIP-qPCR for Cbf1 motifs |
| Sulfur-limited | Broader regulon activation | Genome-wide Met31/Met32 occupancy |
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Mechanistic insight: Constitutive Cbf1 binding primes promoters, while Met31/32 recruitment drives condition-specific MET4 activity .
How to analyze MET4-cofactor stoichiometry in single cells?
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CITE-seq: Couple MET4 antibody with oligonucleotide tags for simultaneous RNA and protein quantification .
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Spatial proteomics: Use multiplexed imaging (CyTOF) with metal-tagged antibodies against MET4, Cbf1, and Met28 .
What strategies mitigate spectral overlap in MET4 multiplex panels?
| Fluorophore | Excitation/Emission (nm) | Compatible Partners |
|---|---|---|
| Pacific Blue | 405/450 | FITC, PE-Cy7 (minimal overlap) |
| AF647 | 640/670 | BV605, PE-Dazzle594 |
Methodological Best Practices
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Genome-wide profiling: Combine ChIP-seq with RNA-seq under Met4 hyperactivation (e.g., GAL1-driven MET4 overexpression) to link binding to transcriptional output .
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Dynamic range: For quantitative Western blotting, use MET4-inducible strains to generate a standard curve (0–90 min induction) .
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Data contradiction analysis: If MET4 binding does not correlate with gene activation, check for post-translational modifications (e.g., ubiquitination status via anti-ubiquitin blots) .
