GSTF7 Antibody
Description
Introduction to GSTF7 Antibody
The GSTF7 antibody is a polyclonal antibody specifically designed to detect glutathione S-transferase F7 (GSTF7), a member of the phi-class glutathione S-transferase (GST) family in Arabidopsis thaliana (Mouse-ear cress). GSTs are multifunctional enzymes involved in detoxification, stress response, and cellular homeostasis. GSTF7 (also designated ATGSTF7 or GST11) is implicated in oxidative stress management and xenobiotic metabolism in plants. This antibody serves as a critical tool for studying GSTF7’s role in plant physiology and stress adaptation .
Gene and Protein Structure
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Protein Class: Phi-class GST (GSTF), which conjugates glutathione to electrophilic substrates .
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Domains: Contains conserved GST-N and GST-C terminal domains typical of GST superfamily members .
Key Features
Production Workflow
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Antigen Preparation: Recombinant GSTF7 expressed in E. coli.
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Immunization: Rabbits immunized with purified GSTF7.
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Purification: IgG fraction isolated via affinity chromatography .
Key Uses
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Stress Response Studies: Detects GSTF7 expression under oxidative stress or herbicide exposure in Arabidopsis .
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Protein Localization: Identifies subcellular distribution in plant tissues via immunofluorescence .
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Metabolic Pathway Analysis: Investigates GSTF7’s role in the mercapturic acid pathway .
Case Study: GSTF7 in Herbicide Resistance
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Finding: Overexpression of GSTF7 in transgenic Arabidopsis correlates with enhanced detoxification of herbicides like atrazine .
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Method: Western Blot analysis using GSTF7 antibody confirmed protein upregulation .
Limitations
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No clinical data available (GSTF7 is plant-specific).
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Limited structural studies compared to mammalian GSTs (e.g., GSTP1) .
Comparative Analysis with Related Antibodies
| Feature | GSTF7 Antibody | Anti-GSTP1 (Human) | General GST Tag Antibodies |
|---|---|---|---|
| Specificity | Plant-specific (Arabidopsis) | Human-specific | Broad (e.g., Schistosoma GST) |
| Applications | Plant stress studies | Cancer research | Recombinant protein detection |
| Commercial Sources | MyBioSource, Agrisera | BD Biosciences, Cell Signaling Tech | Thermo Fisher, Sigma-Aldrich |
Future Directions
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- AtGST11 protein levels are induced by various abiotic stresses including cold, heat, diamide, hydrogen peroxide (H2O2), aluminum, cadmium, copper, and zinc. Notably, the expression of GST11 is completely repressed by heat stress. PMID: 23009097
Q&A
Basic Research Questions
What experimental approaches validate the specificity of GD3G7 antibody for chondroitin sulfate-E (CS-E) in ovarian cancer studies?
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Methodology: Specificity is validated using ELISA competition assays with CS-E and structurally distinct glycosaminoglycans (CS-A, DS, CS-C) to confirm selective binding .
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Immunohistochemical validation: Staining of ovarian adenocarcinoma tissues compared to normal ovaries/cystadenomas (Table 1) .
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Correlation with enzymatic activity: Expression of GalNAc4S-6st (critical for CS-E biosynthesis) in cell lines correlates with GD3G7 reactivity .
Table 1: GD3G7 Staining Patterns in Ovarian Tissues
| Tissue Type | Stromal Staining | Tumor Cell Staining | Basement Membrane Staining |
|---|---|---|---|
| Normal Ovary | None | None | None |
| Cystadenomas | None | None | None |
| Adenocarcinomas | Strong | Occasional Weak | Strong |
How is the GSTF7 promoter used as a bioreporter to screen microbial biocontrol agents in plant studies?
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Reporter system: GSTF7 (a glutathione S-transferase gene) is linked to luciferase (GSTF7:luc) to monitor stress responses in Arabidopsis .
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High-throughput workflow:
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Key application: Identified Streptomyces KB001 as a biocontrol agent against Sclerotinia sclerotiorum via ROS and JA pathway activation .
Advanced Research Questions
How does GD3G7 antibody-mediated CS-E binding influence VEGF interactions in tumor microenvironments?
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Mechanistic insight: CS-E binds VEGF, and GD3G7 competes with VEGF for CS-E binding, suggesting a regulatory role in tumor angiogenesis .
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Functional assays:
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Contradictions: While CS-E is abundant in tumor stroma, its direct role in VEGF signaling requires further in vivo validation .
What transcriptomic insights reveal GSTF7-linked defense mechanisms against necrotrophic fungi?
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RNAseq findings:
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Comparative analysis: Culture filtrate of Streptomyces KB001 induced stronger defense responses than whole microbial treatment, likely due to pre-synthesized metabolites .
Table 2: Top Induced Genes in Arabidopsis Post-KB001 Treatment
| Gene | Function | Fold Change (6 h) | Fold Change (24 h) |
|---|---|---|---|
| GSTF7 | ROS detoxification | 12.5x | 8.3x |
| PR1 | Pathogen response | 9.1x | 6.7x |
| ERF6 | JA signaling | 7.8x | 10.2x |
How are recombinant antibody fragments engineered for improved tumor targeting?
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Design strategies:
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Validation: Competitive ELISAs and in vivo half-life assessments (e.g., chimeric vs. humanized formats) .
What experimental discrepancies arise when analyzing CS-E expression across ovarian cancer subtypes?
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Key矛盾: Endometrioid and serous adenocarcinomas show weak tumor-cell CS-E expression, while mucinous subtypes exhibit stromal-only staining .
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Resolution strategies:
Methodological Recommendations
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For antibody validation: Combine ELISA, immunohistochemistry, and enzymatic activity assays to address off-target binding risks .
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For plant-pathogen studies: Pair GSTF7:luc reporters with RNAseq to map temporal defense pathways .
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For recombinant antibodies: Prioritize phage display libraries with diversity-enhancing mutations to improve affinity .
