ogt-1 Antibody
Description
Introduction to OGT-1 Antibodies
OGT-1 antibodies are specialized immunological reagents designed to detect and study O-linked N-acetylglucosamine transferase (OGT), the sole enzyme responsible for catalyzing O-GlcNAcylation—a post-translational modification critical for regulating protein function, stability, and localization . These antibodies are indispensable tools in research, enabling the analysis of OGT’s role in diverse biological processes, including metabolic regulation, stress responses, cancer progression, and immune modulation .
Applications of OGT-1 Antibodies
OGT-1 antibodies are validated for diverse experimental techniques, as detailed below:
Detection Methods
Key Research Insights
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Hypertonic Stress Response: OGT’s TPR domain (non-catalytic) regulates osmoprotective protein translation in C. elegans and mammals .
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Cancer Immunology: OGT inhibition enhances type I interferon signaling via cGAS-STING activation, synergizing with anti-PD-L1 therapies in colorectal cancer models .
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Novel Shark Antibodies: Single-domain antibodies (e.g., 3F7) enable live-cell imaging and super-resolution microscopy for OGT tracking .
OGT in Hypertonic Stress
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C. elegans Studies: ogt-1 mutants fail to upregulate osmoprotective proteins (e.g., GPDH-1) under hypertonic conditions, despite normal mRNA levels. The TPR domain is essential for this post-transcriptional regulation .
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Conservation: Human OGT partially rescues ogt-1 defects, suggesting evolutionary conservation of non-catalytic OGT functions .
OGT in Cancer and Immunotherapy
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cGAS-STING Activation: OGT inhibition (e.g., via OSMI-1) induces cytosolic DNA accumulation, triggering antitumor immunity and synergizing with anti-PD-L1 therapy in mouse models .
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HCF1 Regulation: OGT-mediated glycosylation of HCF1 is critical for maintaining genomic integrity in colorectal cancer .
Innovations in Antibody Technology
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- A study in *C. elegans* and humans has revealed an evolutionary conserved immunity module involving the nutrient sensor OGT-1. This module establishes a previously unknown connection between nutrient availability and pathogen-specific immune responses. PMID: 25474640
Q&A
Here’s a structured collection of FAQs tailored for academic researchers working with ogt-1 antibodies, organized by complexity and informed by peer-reviewed studies:
How do I validate the specificity of an ogt-1 antibody in my experimental system?
Methodological Answer:
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Knockout/knockdown controls: Use ogt-1 knockout cell lines (e.g., CRISPR-edited models) or siRNA-mediated knockdown to confirm loss of signal in Western blot (WB) or immunofluorescence (IF) .
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Cross-validation: Compare results with orthogonal methods (e.g., RNAi + rescue experiments or mass spectrometry-based O-GlcNAc profiling) .
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Species cross-reactivity: Verify antibody reactivity across species (e.g., human, C. elegans) using recombinant proteins or lysates from validated models .
Example Data Table:
| Validation Method | Expected Outcome | Key Citation |
|---|---|---|
| WB in ogt-1 KO cells | No band at ~117 kDa | |
| IF in C. elegans | Nuclear/cytoplasmic staining |
What are the most robust applications for ogt-1 antibodies in mechanistic studies?
Methodological Answer:
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Chromatin immunoprecipitation (ChIP): Use validated antibodies (e.g., Active Motif 61355) for studying OGT-1’s role in epigenetic regulation (e.g., HCF-1 interaction, histone H2B GlcNAcylation) .
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Subcellular localization: Combine IF with compartment-specific markers (e.g., lamin B1 for nucleus) to resolve OGT-1’s dual nuclear/cytoplasmic roles .
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Functional assays: Pair antibody-based detection with enzymatic inhibition (e.g., OSMI-1) to dissect OGT-1’s catalytic vs. scaffolding roles .
How can I resolve contradictory data on ogt-1’s role in DNA damage versus immune activation?
Methodological Answer:
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Context-dependent analysis:
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Tissue-specific controls: Compare OGT-1 expression levels (WB) across tissues (e.g., pancreas vs. brain) to interpret phenotype variability .
Example Data Comparison:
| Model System | OGT-1 Role | Key Readout | Citation |
|---|---|---|---|
| CRC tumors | Immune evasion | CD8+ T cell infiltration ↓ | |
| C. elegans | Osmoadaptation | GPDH-1 protein stabilization |
What strategies improve ogt-1 antibody performance in low-abundance targets?
Methodological Answer:
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Signal amplification: Use biotinylated shark VNARs (e.g., 3F7 with KD = 53.4 nM) with streptavidin-HRP for enhanced ELISA/WB sensitivity .
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Pre-clearing lysates: Remove high-abundance OGT-1-binding proteins (e.g., HCF-1) via immunodepletion before IP .
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Crosslinking ChIP: Optimize formaldehyde fixation time (e.g., 10 min) to capture transient OGT-1-chromatin interactions .
How do I address cross-reactivity with OGT isoforms or paralogs?
Methodological Answer:
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Epitope mapping: Use antibodies targeting isoform-unique regions (e.g., N-terminal immunogen in Active Motif 61355) .
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Phylogenetic analysis: Test cross-reactivity in species with divergent OGT-1 sequences (e.g., shark vs. human) using recombinant proteins .
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Computational docking: Predict antibody-paratope interactions (e.g., 3F7 binds OGT Ser375/Tyr380) to identify isoform-specific motifs .
Why do some studies report nuclear OGT-1 localization while others emphasize cytoplasmic roles?
Methodological Insights:
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Fixation artifacts: Compare methanol (-20°C) vs. paraformaldehyde (RT) fixation in IF to preserve sublocalization .
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Cell cycle synchronization: OGT-1 translocates to the nucleus during S phase; use FUCCI reporters for phase-specific analysis .
Key Citations for Methodology:
