GLP9 Antibody
Description
Definition and Target Overview
GLP9 Antibody is an immunological reagent designed to detect and bind to the GLP9 protein. While the exact biological role of GLP9 remains unclear, commercial suppliers list it as a target for research applications. Existing products are primarily validated for Arabidopsis thaliana (a model plant organism), suggesting potential relevance in plant biology studies .
Potential Research Applications
Based on technical specifications:
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Plant Biology: GLP9 may play a role in Arabidopsis metabolic or developmental pathways, though mechanistic insights are absent.
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Biomarker Studies: Antibodies could aid in protein expression profiling under stress or genetic modification conditions.
Limitations and Knowledge Gaps
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Target Ambiguity: The term "GLP9" is not widely recognized in major protein databases (e.g., UniProt, NCBI), raising questions about its identity versus homologs like GLP-1 (a well-studied human glucagon-like peptide).
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Species Restriction: Reactivity is confined to Arabidopsis, limiting translational relevance to mammalian systems.
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Validation Deficits: No published figures, functional assays, or cross-reactivity data are available for these products .
Comparison to Related Antibodies
While GLP9 remains obscure, antibodies against GLP-1 (e.g., ab26278 , bsm-0933M ) are extensively validated for diabetes and metabolic research. Unlike GLP9, GLP-1 antibodies:
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Target human/mouse/rat proteins.
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Are used in therapeutic contexts (e.g., blood glucose regulation) .
Future Directions
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Target Identification: Clarify whether GLP9 is a plant-specific protein or a nomenclature variant of established targets.
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Functional Studies: Develop knockout Arabidopsis models to elucidate GLP9’s role.
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Antibody Optimization: Expand validation to include immunohistochemistry and in vivo applications.
Product Specs
Target Background
Q&A
FAQs for GLP9 Antibody Research
What experimental designs are optimal for analyzing GLP9 antibody performance in transient expression systems?
Advanced Research Focus
Transient systems (e.g., plant protoplasts or HEK293 cells) enable rapid testing of antibody-antigen interactions :
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Dose-response assays: Titrate antibody concentrations to establish linear dynamic ranges.
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Co-expression with tagged TFs: Use systems like TARGET (Transient Assay Reporting Genome-wide Effects) to monitor interactions in real time .
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Include translation inhibitors: Cyclohexamide pretreatment isolates direct transcriptional effects from secondary responses .
Key Considerations:
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Transient systems lack stable genomic integration, necessitating repeated transfections for longitudinal studies .
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Pair with ChIP-Seq to map DNA-binding sites of GLP9-associated transcription factors .
How should researchers address discrepancies between computational antibody design predictions and empirical binding data?
Advanced Research Focus
Discrepancies often arise from:
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In silico limitations: RosettaAntibodyDesign (RAbD) may overestimate binding affinity (e.g., predicted ΔG = -40 REU vs. no binding in SPR) .
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Post-translational modifications: Computational models rarely account for glycosylation or phosphorylation .
Resolution Strategy:
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Validate designs using in vitro expression systems (e.g., CHO or HEK cells) .
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Perform alanine scanning mutagenesis to identify critical residues for binding .
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Compare computational models with cryo-EM or X-ray crystallography data .
What methodological frameworks ensure reproducibility of GLP9 antibody-based assays?
Basic Research Focus
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Standardized protocols: Adopt guidelines from the International Working Group on Antibody Validation (IWGAV) .
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Batch-to-batch consistency: Use SDS-PAGE and size-exclusion chromatography to verify antibody purity .
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Cross-lab collaboration: Share positive/negative control samples to harmonize results .
Advanced Research Focus
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Time-course experiments: Measure mRNA/protein levels at multiple timepoints post-perturbation .
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TF fusion systems: Use glucocorticoid receptor (GR) fusions to synchronize GLP9 nuclear translocation .
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Combine ChIP-Seq and RNA-Seq: Identify direct targets via co-occurrence of DNA binding and transcriptional changes .
Data Interpretation:
| Observation | Direct Effect Indicator | Indirect Effect Indicator |
|---|---|---|
| Rapid mRNA change (≤2 hrs) | ✔️ | ❌ |
| Delayed protein expression (≥24 hrs) | ❌ | ✔️ |
What strategies mitigate nonspecific binding in GLP9 antibody applications?
Basic Research Focus
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Pre-adsorption: Incubate antibodies with tissue lysates from non-target species .
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Buffer optimization: Increase salt concentrations (e.g., 150 mM NaCl) to reduce electrostatic interactions .
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Fragment antigen-binding (Fab) use: Replace full-length IgG with Fab fragments to minimize Fc-mediated binding .
Case Study:
A chimeric GLP9 antibody exhibited 40% nonspecific binding in murine brain sections. Humanization reduced off-target interactions by 65% while retaining affinity .
