AAE12 Antibody
Description
Target Overview: ALOX12 (Arachidonate 12-Lipoxygenase)
ALOX12 is a lipid-metabolizing enzyme that catalyzes the oxygenation of polyunsaturated fatty acids (e.g., arachidonic acid) into bioactive lipid mediators like 12-hydroxyeicosatetraenoic acid (12-HETE). It plays roles in:
Mechanistic Roles
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Lipid Mediator Synthesis: Converts arachidonic acid to (12S)-HPETE, a pro-inflammatory and pro-thrombotic mediator .
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Pathological Roles:
Therapeutic Potential
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Preclinical studies suggest ALOX12 inhibition may suppress tumor growth and inflammation .
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AAV-mediated monoclonal antibody delivery shows promise in targeting diseases like HIV and cancer .
Key Studies
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HIV Protection: AAV-delivered ALOX12-targeting immunoadhesins protected macaques from SIV infection .
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Platelet Activation: Neutralizing ALOX12 antibodies reduced thrombus formation in murine models .
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Structural Analysis: Cryo-EM studies of analogous antibodies (e.g., SARS-CoV-2 neutralizing antibodies) highlight the importance of epitope specificity for therapeutic efficacy .
Recommended Dilutions
| Application | Dilution |
|---|---|
| Western Blot | 1:1000 |
| Immunofluorescence | 1:100 |
| IHC-Paraffin | 1:10–50 |
Validation
Comparative Antibody Strategies
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Here’s a structured, research-focused FAQ for the AAE12 antibody, designed for academic researchers. The content synthesizes principles from antibody validation, experimental design, and database utilization, based on published methodologies and best practices :
How can I validate the specificity of AAE12 antibody for my experimental model?
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Method: Perform a multi-step validation workflow:
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Knockout/Knockdown controls: Use CRISPR/Cas9-modified cell lines lacking the AAE12 target protein to confirm absence of signal .
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Orthogonal validation: Compare results with mass spectrometry or RNAi-mediated protein suppression.
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Cross-reactivity checks: Test antibody reactivity against homologous proteins (e.g., paralogs with >80% sequence similarity) using peptide arrays or recombinant proteins .
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What experimental conditions optimize AAE12 antibody performance in immunofluorescence (IF)?
| Parameter | Recommended Condition | Rationale |
|---|---|---|
| Fixation | 4% PFA (15 min) | Preserves epitope integrity |
| Permeabilization | 0.1% Triton X-100 (10 min) | Balances membrane access & structure |
| Antibody dilution | 1:200 (in 1% BSA/PBS) | Minimizes background noise |
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Validation data: Ensure subcellular localization matches databases like AAgAtlas (e.g., nuclear vs. cytoplasmic) .
How do I resolve conflicting AAE12 antibody data across published studies?
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Analysis framework:
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Application-specific validation: Confirm the antibody’s validation status for your method (e.g., IF vs. Western blot) using CiteAb’s citation filters .
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Contextual factors: Compare cell/tissue types, fixation methods, and post-translational modifications.
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Data triangulation: Cross-reference with transcriptomic (RNA-seq) or proteomic (mass spec) datasets to verify expected expression patterns.
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What statistical approaches are robust for quantifying AAE12 expression in heterogeneous samples?
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Recommended workflow:
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Single-cell resolution: Use imaging-based quantitation (e.g., Volocity or ImageJ macros) with background subtraction.
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Normalization: Express AAE12 levels relative to housekeeping proteins (e.g., GAPDH) validated in your system.
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Thresholding: Apply ROC curve analysis to distinguish positive vs. negative signals in clinical specimens .
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Methodological Challenges Table
Key Considerations for Experimental Design
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Lot-to-lot variability: Always include a positive control from prior experiments when using new antibody batches .
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Disease relevance: Cross-check AAE12’s association with autoimmune/cancer pathways in AAgAtlas before mechanistic studies .
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Ethical validation: For preclinical models, ensure antibody cross-reactivity with species orthologs (e.g., murine AAE12) is confirmed .
