At3g47520 Antibody
Description
Introduction to At3g47520 Antibody
The At3g47520 antibody targets the plastidial NAD-dependent malate dehydrogenase enzyme, which facilitates the reversible oxidation of malate to oxaloacetate. This enzyme is essential for photorespiration, redox balance, and metabolite shuttling between chloroplasts and mitochondria . The antibody enables researchers to investigate the protein's expression, localization, and functional roles in Arabidopsis mutants.
Development and Validation
The antibody was generated as part of a large-scale effort to create immunocytochemistry-grade tools for Arabidopsis proteins . Key steps include:
Immunogen Design
Affinity Purification
Crude antisera were affinity-purified using recombinant malate dehydrogenase to enhance specificity. This process increased the detection success rate from 0% (crude sera) to 55% (purified antibodies) .
Validation Methods
Metabolic Pathway Analysis
The antibody has been used to study malate dehydrogenase's role in:
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Citrate cycle: Catalyzing malate/oxaloacetate interconversion .
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Photorespiration: Balancing redox equivalents during high light stress .
Functional Studies in Mutants
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Lethality of Null Alleles: Homozygous At3g47520 CRISPR mutants are embryonic lethal, underscoring the enzyme's essential role .
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Phenotypic Suppression: Partial At3g47520 mutations (e.g., som410, som430) suppress cell death in the mod1 mutant, linked to altered mitochondrial retrograde signaling .
Technical Challenges and Solutions
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Purification and characterization of the plastid-localized NAD-dependent malate dehydrogenase from Arabidopsis thaliana PMID: 26095832
- The critical role of plastidial NAD-specific MDH isoform in Arabidopsis (Arabidopsis thaliana) plants has been reported. PMID: 24453164
- Arabidopsis thaliana mutants lacking the NADP-malate dehydrogenase have lost the reversible inactivation of catalase activity and the increase in H2O2 levels when exposed to high light. PMID: 24591715
- Plastid-localized NAD-dependent malate dehydrogenase is crucial for energy homeostasis in developing Arabidopsis thaliana seeds PMID: 24198233
- mMDH plays a role in maximizing photorespiratory rate. PMID: 20876337
