ABCG9 Antibody
Description
Functional Role of ABCG9
ABCG9 is a half-type ATP-binding cassette (ABC) transporter that forms functional dimers to mediate substrate translocation across membranes. Key roles include:
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Pollen Development: In Arabidopsis thaliana, ABCG9 partners with ABCG31 to regulate pollen viability by maintaining lipid and sterol homeostasis. Knockout mutants (abcg9 abcg31) exhibit ~50% pollen lethality, rescued by ABCG9 complementation .
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Vascular Patterning: ABCG9, ABCG11, and ABCG14 are essential for phloem development, likely by transporting sterols or lipids required for vascular cell differentiation .
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Viral Defense: In the insect vector Laodelphax striatellus, ABCG9 suppresses rice black-streaked dwarf virus (RBSDV) infection. Knockdown of LsABCG9 increases viral load, implicating it in antiviral responses .
ABCG9 Antibody Applications
While commercial ABCG9 antibodies are not explicitly detailed in the provided sources, research-grade tools include:
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GFP-Tagged Constructs: Arabidopsis studies used ABCG9 pro:sGFP:ABCG9 transgenes for subcellular localization, confirming plasma membrane targeting in tapetal cells .
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RNA Interference (RNAi): LsABCG9 knockdown in insects validated its role in viral resistance .
Table 1: Key Research Applications of ABCG9-Specific Tools
Technical Considerations for Antibody Use
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Epitope Design: Antigenic regions should avoid conserved ATP-binding domains to ensure specificity.
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Validation: Include controls such as knockout mutants (e.g., abcg9 abcg31) or RNAi-treated samples to confirm antibody specificity .
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Cross-Reactivity: ABCG9 shares homology with other ABCG transporters; rigorous testing across species is critical .
Future Research Directions
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Substrate Identification: Mass spectrometry or radiolabeled assays to define transported molecules.
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Therapeutic Potential: Modulating ABCG9 in crop plants or insect vectors could enhance disease resistance .
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Human Orthologs: Investigating ABCG9 homologs (e.g., ABCB9) in lipid disorders or drug resistance .
Product Specs
Target Background
Gene References and Functions:
- ABCG9, ABCG11, and ABCG14 are expressed in the plant vascular system. Mutations in these genes result in impaired plant growth and disrupted vascular patterning. (ABCG9, PMID: 24112720)
Q&A
Here’s a structured FAQ for researchers working with ABCG9 antibodies, informed by principles of antibody validation, experimental design, and reproducibility from peer-reviewed methodologies:
Advanced Research Questions
How do I resolve contradictions between ABCG9 antibody performance in Western blot vs. IHC?
| Factor | Western Blot | IHC |
|---|---|---|
| Antigen presentation | Denatured, linear epitopes | Native, conformational |
| Common issues | Cross-reactivity | Tissue fixation artifacts |
| Solutions | Validate with knockout | Optimize antigen retrieval |
What structural features of ABCG9 influence antibody-antigen interactions?
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ABCG9’s transmembrane domains may require antibodies targeting extracellular loops for live-cell applications .
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Epitope mapping via alanine scanning or phage display can identify critical binding regions .
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Consider iodoacetamide-treated antigens to stabilize disulfide bonds and improve antibody recognition .
How can I assess ABCG9 antibody cross-reactivity with homologous proteins (e.g., ABCG8)?
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Perform BLAST analysis of the immunogen sequence to identify shared regions.
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Test antibody binding against recombinant ABCG8 in ELISA or surface plasmon resonance (SPR) .
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Use immunoprecipitation-mass spectrometry to identify off-target proteins .
Methodological Best Practices
What statistical approaches improve reproducibility in ABCG9 studies?
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Blinded analysis: Separate experimental and data-analysis teams .
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Power calculations: Use pilot data to determine sample sizes (e.g., ≥3 biological replicates) .
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Report antibody identifiers (RRID, catalog number, lot) and validation data in publications .
How do I design a study to investigate ABCG9’s role in disease mechanisms?
| Stage | Approach | Validation |
|---|---|---|
| Hypothesis | Link ABCG9 to lipid transport pathways | Literature mining (e.g., OMIM) |
| In vitro | siRNA knockdown in relevant cell lines | qPCR + functional assays |
| In vivo | Conditional knockout models | Phenotypic + metabolomic profiling |
Data Interpretation Challenges
How to troubleshoot inconsistent ABCG9 expression data across studies?
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Technical variables: Compare fixation methods (e.g., paraformaldehyde vs. methanol) .
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Biological variables: Account for circadian rhythms or hormonal influences on ABCG9 .
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Antibody validation: Use CiteAb to identify citations supporting antibody performance .
