CML35 Antibody
Description
CSL362 (Anti-CD123)
CSL362 is a humanized monoclonal antibody targeting the interleukin-3 receptor α subunit (CD123), a marker elevated in CML leukemic stem and progenitor cells (LSPCs) compared to healthy cells .
Mechanism of Action:
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Induces antibody-dependent cell-mediated cytotoxicity (ADCC) via natural killer (NK) cells.
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Blocks IL-3–mediated rescue of tyrosine kinase inhibitor (TKI)-induced cell death .
Key Findings:
Clinical Relevance:
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Proposed for consolidation therapy in CP-CML to sustain TKI-induced molecular remission .
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Preferential targeting of CD123+ granulocyte-macrophage progenitors in BC-CML .
4D7 Antibody
4D7 is a murine monoclonal antibody engineered to target the mutant calreticulin (CALR) neoepitope in myelofibrosis, a condition overlapping with CML progression .
Function:
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Inhibits thrombopoietin (TPO)-independent signaling in mutant CALR cells.
Preclinical Data:
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Reduced colony formation in mutant CALR cells by 95% in vitro .
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Improved survival in murine models with ruxolitinib-resistant engraftment .
Anti-CD35 (EP197) in Diagnostic Pathology
CD35 (complement receptor 1) is a glycoprotein expressed on follicular dendritic cells and mature B cells. The EP197 clone (Cell Marque) is used diagnostically to label lymphoproliferative disorders .
Applications:
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Identifies follicular dendritic cell tumors.
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Reactivity: Paraffin-embedded tissues at dilutions of 1:25–1:100 .
Anti-Pleiotrophin Antibody
A preclinical anti-pleiotrophin antibody demonstrated synergistic efficacy with imatinib, eradicating CML xenografts in mice .
Mechanism:
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Targets pleiotrophin, a protein overexpressed in CML stem cells.
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Combined with TKIs, it reduced leukemic engraftment by >90% .
T Cell Receptor Mimic (TCRm) Antibodies
TCRm antibodies targeting peptide-MHC complexes (e.g., WT1, PRAME) are under investigation for CMR maintenance .
Design Considerations:
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Target specificity for cancer vs. normal cells.
Comparative Efficacy of Selected Antibodies
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Basic Research Questions
How to validate specificity of CML35 antibodies in Arabidopsis thaliana studies?
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Perform immunoblotting against recombinant CML35 protein and negative control samples (e.g., cml35 knockout mutants) to confirm target specificity .
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Combine with mass spectrometry to identify off-target binding partners in wild-type extracts.
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Validate cross-reactivity risks by testing against other calmodulin-like proteins (e.g., CML1, CML34) using ELISA .
What experimental controls are critical for CML35 antibody-based assays?
Which methods optimize CML35 detection in Western blotting?
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Use Tris-Tricine gels for improved resolution of low-molecular-weight calcium-binding proteins .
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Pre-block membranes with 5% nonfat milk + 0.1% Tween-20 to reduce background noise.
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Titrate antibody concentrations (start with 1:1,000 dilution) to balance signal-to-noise ratios .
Advanced Research Questions
How to resolve contradictory subcellular localization data for CML35?
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Compare fixation protocols: Paraformaldehyde-based methods may alter calcium-dependent epitope accessibility vs. cryopreservation .
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Pair immunofluorescence with fluorescently tagged CML35 transgenic lines for spatial validation.
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Account for dynamic calcium signaling by sampling tissues at multiple developmental stages.
What strategies identify CML35 interaction partners in calcium signaling networks?
| Approach | Strengths | Limitations |
|---|---|---|
| Co-IP/MS | Identifies direct/indirect interactors | Requires high antibody affinity |
| FRET-FLIM | Quantifies real-time interactions in planta | Technically challenging |
| Yeast two-hybrid | Maps binary interactions | Misses calcium-dependent complexes |
How to differentiate CML35 functional roles across stress responses?
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Develop conditional knockdown lines using CRISPR/dCas9 systems to avoid pleiotropic effects.
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Pair transcriptomic data (RNA-seq under stress) with CML35 protein abundance assays .
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Test functional redundancy by creating higher-order mutants with paralogs (e.g., cml34/cml35 double mutants).
Methodological Considerations
7. Validating antibody performance in plant species beyond Arabidopsis:
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Perform phylogenetic alignment of CML35 homologs to assess epitope conservation.
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Test cross-reactivity with protein extracts from closely related Brassicaceae species (e.g., Brassica napus).
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Quantify binding affinity via surface plasmon resonance if commercial kits are unavailable.
8. Addressing batch-to-batch variability in polyclonal CML35 antibodies:
