NDA1 Antibody
Description
Background and Target Proteins
NDA1 is a component of the NDA-type alternative mitochondrial NAD(P)H dehydrogenases in plants, which bypass the conventional electron transport chain (ETC) to regulate redox balance. These enzymes are critical for stress responses and mitochondrial function.
Antibody Development and Specificity
To study NDA1 function, researchers developed affinity-purified rabbit polyclonal antibodies targeting peptide sequences from Arabidopsis thaliana NDA1 and NDA2. These antibodies were validated using recombinant proteins and Western blotting:
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NDA1 Antibody: Detected a ~48 kDa band in wild-type (WT) mitochondria but showed minimal signal in RNAi-suppressed lines.
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NDA2 Antibody: Detected a ~48 kDa band in WT, with reduced intensity in RNAi lines.
| Parameter | WT | RNAi Line 17.11 | RNAi Line 26.6 |
|---|---|---|---|
| NDA1 mRNA Levels | 100% | 9–36% | 9–36% |
| NDA1 Protein Signal | Detectable | Barely Detectable | Barely Detectable |
| NDA2 mRNA Levels | 100% | 56–78% | 56–78% |
| NDA2 Protein Signal | Detectable | ~50% of WT | ~50% of WT |
Data adapted from suppression studies in A. thaliana .
Functional Insights
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RNAi Suppression: Silencing NDA1/2 reduced mitochondrial NAD(P)H dehydrogenase activity, highlighting their role in redox regulation.
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Protein Localization: NDA1 and NDA2 were localized to mitochondria, with no cross-reactivity to peroxisomal or glyoxysomal isoforms.
Role in DNA Replication
Nda1 (and Nda4) in Schizosaccharomyces pombe are MCM (Mini-Chromosome Maintenance) homologs, essential for initiating DNA replication. Unlike budding yeast MCMs, their localization does not cycle dynamically.
Antibody Development and Localization
Antibodies against Nda1 and Nda4 were raised to study their nuclear dynamics:
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Nda1 Antibody: Detected a 115 kDa protein in nuclear fractions.
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Nda4 Antibody: Detected an 80 kDa protein, co-eluting with Nda1 in affinity chromatography.
| Cell Cycle Phase | Nda1 Localization | Nda4 Localization |
|---|---|---|
| G1 | Nuclear | Nuclear |
| S | Nuclear | Nuclear |
| G2 | Nuclear | Nuclear |
Data from immunolocalization studies in fission yeast .
Functional Insights
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Nuclear Retention: Both proteins remain nuclear throughout the cell cycle, contrasting with budding yeast MCMs.
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Complex Formation: Nda1 and Nda4 form a stable complex, critical for replication licensing.
Comparative Analysis of NDA1 Antibodies
Challenges and Future Directions
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Specificity Concerns: Cross-reactivity with homologs (e.g., NDA2 in plants) requires stringent validation.
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Functional Studies: Linking NDA1/2 activity to stress responses (plants) or replication fidelity (yeast) remains an open area.
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Therapeutic Potential: While not yet explored, antibodies against mitochondrial NDA1 could inform biotechnological strategies for crop resilience.
This synthesis underscores the versatility of NDA1 antibodies in elucidating conserved and divergent biological processes. Further studies integrating proteomics and functional genomics will deepen their utility in research and applied biology.
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
