CXE8 Antibody
Description
Definition and Biological Role of CXCL8
CXCL8 (IL-8) is a proinflammatory chemokine belonging to the CXC family, primarily involved in neutrophil recruitment and angiogenesis . It binds to receptors CXCR1 and CXCR2, activating pathways that promote inflammation, tumor progression, and immune modulation .
CXCL8 Antibody Characteristics
CXCL8 antibodies are tools for detecting or neutralizing CXCL8 in research and clinical settings. Key features include:
3.1. Diagnostic Biomarker Development
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Esophageal Squamous Cell Carcinoma (ESCC): Anti-CXCL8 autoantibodies showed diagnostic potential with AUC values of 0.713–0.751 in distinguishing ESCC patients from controls .
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Table: Diagnostic Performance of Anti-CXCL8 Autoantibody in ESCC
Cohort AUC Sensitivity Specificity Verification Set 0.713 35.7% 82.9% Validation Set 0.751 47.1% 77.9%
3.2. Therapeutic Neutralization
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Acute Lung Injury (ALI): Humanized anti-CXCL8 antibodies reduced neutrophil infiltration and lung damage in rabbit models .
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Cancer Therapy: CXCL8 blockade enhanced anti-tumor immunity in preclinical studies, particularly when combined with immune checkpoint inhibitors .
Mechanisms of Action
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Neutralization: Antibodies like MAB208 inhibit CXCL8-induced chemotaxis (ND₅₀: 0.08–0.4 µg/mL) .
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Epitope Specificity: Most antibodies target the N-terminal ELR motif (e.g., MAB208), critical for CXCR1/2 binding .
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Post-Translational Regulation: Antibodies detect truncated or citrullinated CXCL8 forms, which alter bioactivity .
Clinical Implications
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Biomarker Potential: Elevated serum CXCL8 correlates with poor prognosis in cancers (e.g., gastric, lung) .
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Therapeutic Limitations: Rodents lack CXCL8 orthologs, complicating translational studies .
Key Research Findings
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Structural Insights: CXCL8 forms homodimers or heterodimers with CXCL4, influencing receptor binding .
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Autoantibody Role: Anti-CXCL8 autoantibodies may stabilize CXCL8 in serum, enhancing diagnostic utility .
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Dual Roles in Cancer: CXCL8 promotes angiogenesis via VEGF induction while suppressing anti-tumor immunity .
Future Directions
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Here’s a structured FAQ collection for researchers working with CXE8 antibodies, derived from current scientific literature and technical data:
How do I validate the specificity of a CXE8 antibody for Arabidopsis thaliana studies?
Methodological Answer:
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Perform immunoblotting using recombinant CXE8 protein (36.4 kDa) and compare with plant tissue lysates. Include negative controls (e.g., cxe8 knockout mutants) to confirm absence of nonspecific binding .
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Use competitive ELISA with pre-incubation of the antibody against recombinant CXE8 to block antigen-binding activity .
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Validate with immunohistochemistry on Arabidopsis tissue sections, comparing staining patterns to known CXE8 expression profiles (e.g., root or leaf tissues) .
What are the recommended applications for CXE8 antibodies in plant biology?
Methodological Answer:
| Application | Protocol Notes | Key Controls |
|---|---|---|
| Western Blot | Use 12% SDS-PAGE; reduce samples with β-mercaptoethanol | Include recombinant CXE8 and cxe8 mutant lysates |
| ELISA | Coat plates with 0.5–1 µg/mL recombinant CXE8 | Block with 5% BSA; test serial antibody dilutions |
| Immunoprecipitation | Crosslink antibody to Protein A/G beads | Pre-clear lysates with empty beads |
How should I design experiments to study CXE8’s role in carboxylesterase-mediated stress responses?
Methodological Answer:
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Hypothesis-driven approach: Compare oxidative stress markers (e.g., H₂O₂ levels) in wild-type vs. cxe8 mutants under drought or pathogen exposure.
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Multi-omics integration: Pair antibody-based CXE8 localization (immunofluorescence) with transcriptomic data to identify co-regulated pathways .
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Functional complementation: Express epitope-tagged CXE8 in mutants and track rescue phenotypes using antibody-based detection .
How do I resolve contradictions in CXE8 antibody reactivity across plant species?
Methodological Answer:
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Cross-reactivity testing: Screen the antibody against carboxylesterases from closely related species (e.g., Brassica napus) via immunoblot .
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Epitope mapping: Identify the antibody’s binding region using peptide arrays or alanine scanning mutagenesis of recombinant CXE8 .
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Structural alignment: Compare the target epitope’s sequence (e.g., residues 120–150 in CXE8) with homologs using tools like CLUSTAL Omega .
What statistical methods are appropriate for quantifying CXE8 expression variability in mutant lines?
Methodological Answer:
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Use mixed-effects models to account for batch effects in Western blot/ELISA data.
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Apply ANOVA with Tukey’s post-hoc test for multi-group comparisons (e.g., WT vs. heterozygous vs. homozygous mutants).
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For imaging data (e.g., fluorescence intensity), employ nonparametric tests (Mann-Whitney U) if normality assumptions fail .
Why does my CXE8 antibody produce high background in Arabidopsis root sections?
Methodological Answer:
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Pre-absorption: Incubate the antibody with fixed cxe8 mutant tissue lysates to remove nonspecific binders .
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Optimize fixation: Reduce paraformaldehyde concentration (<4%) to preserve epitope accessibility .
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Signal amplification: Use tyramide-based systems (e.g., TSA) to enhance specificity in low-abundance targets .
How to distinguish between CXE8-specific signals and cross-reactive carboxylesterases?
| Validation Strategy | Outcome Interpretation |
|---|---|
| Knockout validation | Loss of band/staining in mutants confirms specificity |
| Peptide competition | ≥80% signal reduction indicates epitope specificity |
| Orthogonal assay (e.g., enzymatic activity) | Correlation between antibody signal and hydrolase activity strengthens validity |
