CRCK2 Antibody
Description
Crk II (C-terminal region) Antibody
Crk II is an adaptor protein critical for intracellular signaling, particularly downstream of tyrosine kinases. The Crk II (C-terminal region) Antibody (e.g., monoclonal antibody M332) targets the C-terminal domain of Crk II, which regulates interactions with SH2 and SH3 domain-containing proteins .
Table 1: Crk II Antibody Applications
CXCR2-Targeting Antibodies
CXCR2 is a chemokine receptor pivotal in neutrophil recruitment. Antibodies targeting CXCR2’s N-terminal region (e.g., the picomolar-affinity antibody described in sources ) block IL-8 binding, inhibiting neutrophil chemotaxis and inflammation.
Mechanistic and Therapeutic Insights
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Epitope Specificity: Binds to the N-terminal region of CXCR2, competing with IL-8 and disrupting ligand-receptor interactions .
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Preclinical Efficacy:
Table 2: CXCR2 Antibody Performance in Disease Models
Comparative Analysis of Antibody Strategies
While Crk II antibodies focus on intracellular signaling modulation, CXCR2 antibodies target extracellular receptor-ligand interactions. Both strategies highlight the importance of domain-specific targeting:
| Parameter | Crk II Antibody | CXCR2 Antibody |
|---|---|---|
| Target Localization | Intracellular adaptor protein | Cell-surface chemokine receptor |
| Therapeutic Scope | Cancer, developmental disorders | Inflammatory diseases, autoimmune conditions |
| Mechanism | Disrupts SH2/SH3-mediated protein binding | Blocks IL-8 binding and neutrophil migration |
Research Limitations and Future Directions
Product Specs
Components: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Here’s a structured FAQ collection for academic research on CXCR2 antibodies, synthesized from peer-reviewed studies and methodological data:
Basic Research Questions
What are the primary functional assays for validating CXCR2 antibody efficacy in neutrophil studies?
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Methodology:
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Chemotaxis inhibition assays: Measure neutrophil migration toward IL-8/CXCL1 using Boyden chambers or microfluidic devices. Antibodies blocking ≥80% migration at 10 nM are considered potent .
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Receptor occupancy assays: Use flow cytometry to quantify CXCR2 surface expression pre/post antibody treatment .
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In vivo validation: Employ CXCR2-humanized mice with induced inflammation (e.g., LPS-induced lung injury) and track neutrophil depletion via histopathology .
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How do researchers confirm CXCR2 antibody specificity?
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Approach:
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Competition binding assays: Test against recombinant CXCR1 (closest homolog) using Biolayer Interferometry (BLI) or Surface Plasmon Resonance (SPR) .
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Epitope mapping: Combine alanine scanning mutagenesis (e.g., N-terminal domain residues like Asp10, Leu15) with antibody binding kinetics .
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Knockout validation: Compare binding in WT vs. CXCR2-KO cell lines via flow cytometry .
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Advanced Research Challenges
How to resolve discrepancies in CXCR2 antibody neutralization potency across disease models?
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Analysis framework:
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Recommendation: Normalize dosing to receptor occupancy (e.g., ≥90% saturation for RA) rather than fixed concentrations .
What computational methods improve CXCR2 antibody affinity while retaining epitope specificity?
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In silico workflow:
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Molecular dynamics: Simulate antibody-CXCR2 interactions (e.g., binding energy < -50 kcal/mol for lead candidates) .
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Directed evolution: Use RosettaAntibodyDesign to optimize CDR-H3 regions, prioritizing ΔΔG < -3.0 kcal/mol variants .
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Validation: Confirm improved variants via BLI (e.g., KD < 100 pM from initial 2 nM) .
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Methodological Comparisons
How do antibody characterization platforms compare for CXCR2 studies?
| Platform | Throughput | KD Range | Key Advantage | Limitation |
|---|---|---|---|---|
| BLI (Octet) | High (96-well) | 1 pM – 100 nM | Real-time kinetics in crude lysates | Limited temperature control |
| SPR (Biacore) | Medium | 10 pM – 1 μM | Gold-standard for regulatory work | High sample consumption |
| Flow cytometry | High | Qualitative | Native receptor conformation | No kinetic parameters |
Contradiction Management
Why do some CXCR2 antibodies show pro-inflammatory effects at low doses?
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Hypothesis testing:
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Check FcγR binding: Use SPR to measure Fc-C1q interaction (affinity > 10^6 M^-1 may cause ADCP) .
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Titrate agonism: Perform calcium flux assays with 0.1–100 nM antibody. EC50 < 1 nM suggests intrinsic signaling .
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In vivo dose-ranging: Compare 0.1 mg/kg vs. 10 mg/kg in zymosan-induced peritonitis; optimal efficacy typically occurs at 1–5 mg/kg .
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