DREB1C Antibody
Description
Biological Role of DREB1C
DREB1C acts as a master regulator of stress adaptation by binding to dehydration-responsive elements (DRE/CRT) in promoter regions of target genes. Key functions include:
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Nitrogen Use Efficiency (NUE): Overexpression of DREB1C in rice enhances 15N uptake, photosynthetic efficiency (via increased RuBisCO content), and nitrogen remobilization to grains .
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Cold Tolerance: In Arabidopsis, DREB1C interacts with circadian clock proteins (CCA1/LHY, RVE4/RVE8) to activate cold-responsive genes like DREB1A and COR15A .
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Drought and Salt Tolerance: Transgenic wheat overexpressing soybean GmDREB1 (a homolog) shows improved root biomass, melatonin biosynthesis, and yield under drought .
Applications of DREB1C Antibodies
While the provided sources do not explicitly detail commercial DREB1C antibodies, their inferred applications include:
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Protein Localization: Immunolocalization to study tissue-specific expression under stress.
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Western Blotting: Quantifying DREB1C levels in transgenic vs. wild-type plants .
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Chromatin Immunoprecipitation (ChIP): Identifying direct DNA targets (e.g., NRT2.4, FTL3) .
Table 1: DREB1C-Regulated Genes and Phenotypic Outcomes
Table 2: Cross-Species Validation of DREB1C Function
Mechanistic Insights
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Transcriptional Regulation: DREB1C binds to DRE/CRT motifs in promoters of stress-responsive genes (e.g., DREB1A, RD29A) .
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Post-Translational Modifications: Cold stress induces degradation of repressors (CCA1/LHY) and stabilizes activators (RVE4/RVE8), enabling DREB1C-mediated gene activation .
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Synergy With Hormones: Overexpression correlates with elevated melatonin and proline, mitigating oxidative damage .
Challenges and Future Directions
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Field Validation: Large-scale trials are needed to confirm yield benefits in elite crop varieties .
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Antibody Specificity: Current studies rely on transgenic tags (e.g., GFP fusion) ; isoform-specific antibodies could refine functional studies.
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CRISPR Applications: Knockout mutants (OsDREB1C/E/G) reveal functional redundancy in rice abiotic stress responses .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- This study reveals the essential functions of C-repeat binding factors (CBF) in chilling stress response and cold acclimation, and defines a set of genes as the CBF regulon. PMID: 27353960
- CBFs play an important role in the trade-off between cold tolerance and plant growth through the precise regulation of COR genes within the complex transcriptional network. PMID: 28009483
- The three CBF genes together are required for cold acclimation and freezing tolerance. PMID: 27252305
- Data indicate that the C-repeat binding factor (CBF) locus encompasses three genes - CBF1, CBF2, and CBF3 (AT4G25480) - which are induced by low temperature and encode transcription factors. PMID: 26369909
- A major locus harboring three cold-responsive transcription factor genes, including CBF1, was identified. PMID: 23721132
- Studies indicate that DREB1A (CBF3), DREB1B (CBF1), and DREB1C (CBF2) play a significant role in enhancing stress tolerance. PMID: 23271026
- Transcriptional changes in cell wall metabolism-related components under overexpression of CBF2, a key transcription factor for freezing tolerance, were identified. PMID: 21611181
- Overexpression of CBF2 in Arabidopsis suppressed the responsiveness of leaf tissues to ethylene compared to wild-type plants, leading to significantly delayed senescence and chlorophyll degradation. PMID: 20636906
- These findings indicate that overexpression of CBF2 not only increases frost tolerance but also affects other developmental processes. PMID: 19854800
- SOC1 directly represses the expression of CBF2 genes. PMID: 19825833
- PIF7 functions as a transcriptional repressor for DREB1C expression, and its activity is regulated by PIF7-interacting factors TIMING OF CAB EXPRESSION1 and Phytochrome B. PMID: 19837816
- We explored the regulation of CBF1-3 by the circadian clock. PMID: 15728337
- The low freezing tolerance of FTQ4-Cvi (FREEZING TOLERANCE QTL 4) alleles was associated with a deletion of the promoter region of Cvi CBF2, and with low RNA expression of CBF2 and of several CBF target genes. PMID: 16244146
- CBF1 and CBF3, but not CBF2, have a concerted additive effect to induce the entire CBF regulon and the complete development of cold acclimation. PMID: 18093929
- Important evolutionary changes in CBF1, -2, and -3 may have primarily occurred at the level of gene regulation as well as in protein function. PMID: 18990244
Q&A
How to validate the specificity of DREB1C antibodies in plant stress studies?
Methodological Answer:
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Western Blot with Knockout Mutants: Use Arabidopsis thaliana mutants lacking functional DREB1C (e.g., dreb1c T-DNA insertion lines) to confirm antibody specificity. A lack of signal in mutants confirms specificity .
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Immunofluorescence with Competitive Peptide Blocking: Pre-incubate the antibody with a synthetic peptide matching the DREB1C epitope (e.g., residues within the AP2/ERF domain). Loss of signal indicates specificity .
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Cross-Reactivity Screening: Test against recombinant proteins of closely related DREB subfamily members (e.g., DREB1A, DREB2A) to rule out off-target binding .
What experimental controls are critical for ChIP-qPCR studies using DREB1C antibodies?
Advanced Considerations:
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No-Antibody Control: Rule out nonspecific DNA pull-down during chromatin immunoprecipitation.
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Isotype Control Antibody: Use a nonspecific IgG to assess baseline noise .
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Mutant Promoter Regions: Include primers targeting genomic regions lacking DREB1C-binding cis-elements (e.g., mutated EE motifs) to validate binding specificity .
How to resolve contradictory data on DREB1C expression levels under cold stress?
Data Contradiction Analysis:
What structural features of DREB1C should guide antibody design for functional studies?
Key Epitope Selection Criteria:
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AP2/ERF Domain: Target residues critical for DNA binding (e.g., Val14 in the β-sheet structure) .
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Nuclear Localization Signal (NLS): Antibodies against the NLS (e.g., PKK/RPAGRxKFxETRHP) can disrupt nuclear import, aiding functional assays .
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Post-Translational Modifications: Use antibodies recognizing phosphorylation sites (e.g., Ser/Thr residues) to study stress-induced activation .
How to optimize DREB1C antibody concentrations for EMSA and ELISA?
Titration Protocol:
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EMSA: Start with 0.5–2 µg of purified DREB1C protein and titrate antibody (0.1–1.0 µg/µL) to observe supershift without nonspecific probe binding .
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ELISA: Coat plates with 100 ng recombinant DREB1C. Test antibody dilutions (1:500–1:10,000) to achieve a signal-to-noise ratio >3:1 .
What are the pitfalls in interpreting DREB1C subcellular localization data?
Technical Challenges:
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Cross-Reactivity with RVEs: RVE4/RVE8 proteins bind overlapping promoter regions and may colocalize with DREB1C, requiring co-staining with RVE-specific antibodies .
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Fixation Artifacts: Over-fixation with paraformaldehyde (>4%) masks epitopes. Optimize fixation time (10–15 min) for root or leaf tissues .
How to integrate DREB1C antibody data with transcriptomic datasets?
Multi-Omics Workflow:
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Correlate Protein-DNA Binding (ChIP-seq): Use anti-DREB1C ChIP-seq peaks to validate differentially expressed genes in RNA-seq data (e.g., COR15A, RD29A) .
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Leverage Public Repositories: Cross-reference with datasets from Phytozome or TAIR for conserved target motifs .
Why do DREB1C antibody signals vary in diurnal vs. stress conditions?
Mechanistic Insight:
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Circadian Regulation: CCA1/LHY transcription factors repress DREB1C under non-stress conditions but degrade under cold stress, enabling RVE4/RVE8-mediated activation .
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Protein Turnover: Cold-induced proteasomal degradation of repressors (e.g., ICE1) alters DREB1C stability. Use MG132 (proteasome inhibitor) to stabilize signals .
