At2g31350 Antibody
Description
Introduction
The At2g31350 Antibody (product code CSB-PA874425XA01DOA) is a research-grade antibody targeting the protein encoded by the AT2G31350 gene in Arabidopsis thaliana (thale cress). This gene, also known as GLX2-5, encodes a mitochondrial glyoxalase 2 (Glx II) critical for detoxifying reactive carbonyl compounds like methylglyoxal. The antibody is primarily used in biochemical studies to investigate protein expression, localization, and functional roles in plant stress responses.
Gene Information
The AT2G31350 gene encodes a mitochondrial glyoxalase 2, part of the glyoxalase pathway responsible for detoxifying methylglyoxal—a toxic byproduct of glycolysis. Key features include:
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Metal Center: Accommodates diverse metal ions, with Fe(III)Zn(II) as the predominant configuration .
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Function: Converts methylglyoxal to D-lactate via glutathione-dependent catalysis, mitigating oxidative stress .
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Localization: Mitochondria, consistent with its role in detoxifying reactive carbonyls in cellular energy hubs .
| Gene Detail | Description |
|---|---|
| DB Identifier | AT2G31350 |
| Symbol | GLX2-5 |
| Pathway | Glyoxalase pathway (KEGG: K01069) |
| Ortholog Groups | KEGG Orthology (KO) |
Research Applications
While direct studies using the At2g31350 Antibody are not explicitly documented in the provided sources, its potential applications align with the gene’s known functions:
Stress Response Analysis
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Oxidative Stress: Investigate Glx II expression under methylglyoxal-induced stress or abiotic conditions.
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Protein Stability: Track post-translational modifications (e.g., metal center dynamics) under stress via Western blotting or immunoprecipitation.
Functional Studies
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Subcellular Localization: Confirm mitochondrial targeting using immunofluorescence or organelle fractionation.
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Protein Interactions: Identify binding partners in the glyoxalase pathway (e.g., glyoxalase I or glutathione) via co-immunoprecipitation.
Comparative Biochemistry
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Metal Ion Specificity: Compare Fe(III)Zn(II) versus other metal configurations (e.g., Fe(III)Fe(III)) in catalysis efficiency.
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Evolutionary Analysis: Cross-reactivity tests with orthologs in other plant species to study glyoxalase evolution.
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Basic Research Questions
How to validate At2g31350 antibody specificity in Arabidopsis mitochondrial protein extracts?
Perform sequential validation using:
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Knockout mutants (e.g., T-DNA insertion lines) as negative controls
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Parallel detection with anti-HA tagged recombinant At2g31350 ( )
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2D gel electrophoresis to confirm predicted molecular weight (35-40 kDa) and isoelectric point (pI 6.2)
What experimental controls are essential when measuring GLX2-5 protein levels under stress conditions?
Include:
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Tissue-specific extraction controls (root vs. leaf mitochondria)
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Co-analysis with glyoxalase I activity using spectrophotometric assays ( )
How to optimize Western blot conditions for detecting low-abundance At2g31350?
Use:
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15-20 μg mitochondrial protein per lane
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Prolonged transfer times (2 hr at 100V) for 35-40 kDa proteins
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Chemiluminescent substrates with high sensitivity (e.g., ECL Prime)
Advanced Research Challenges
How to resolve contradictory Western blot signals between different Arabidopsis ecotypes?
What methodology combines At2g31350 immunodetection with functional glyoxalase pathway analysis?
Integrated workflow:
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Quantify MG-H1 adducts using anti-MG-H1 antibodies (STA 011) ( )
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Measure GLO2 enzymatic activity via spectrophotometric SLG hydrolysis assay
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Correlate protein levels with:
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Glutathione redox status (GSH:GSSG ratio)
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ROS markers (H2O2, O2⁻)
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How to design tissue-specific studies of At2g31350 in root development?
Implement:
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Laser-capture microdissection of root maturation zones
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Subcellular fractionation validation (mitochondrial vs cytosolic)
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Spatial expression mapping using ProAt2g31350:GUS lines
Technical Optimization
What causes variable antibody performance in different Arabidopsis organs?
Critical factors:
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Tissue-specific protease activity (add 2 mM PMSF in extraction buffer)
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Organ-specific glycosylation patterns (use PNGase F treatment)
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Mitochondrial integrity (verify via cytochrome c oxidase assay)
How to integrate At2g31350 antibody data with metabolomic profiling?
Multi-omics strategy:
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Parallel measurement of:
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Multivariate analysis using:
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Partial Least Squares Regression
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Pathway enrichment in MapMan
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Biological Interpretation
How to distinguish developmental vs stress-induced regulation of GLX2-5?
Experimental design:
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Time-course analysis across developmental stages
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Comparative treatments:
Condition MG Level Expected ΔGLX2-5 Control 0.5 μM Baseline NaCl 150mM 2.8 μM +300% Methylviologen 10μM 4.1 μM +450%
What evidence suggests crosstalk between glyoxalase and other detox systems?
Key findings:
