CAMTA2 Antibody
Description
Introduction to CAMTA2 and CAMTA2 Antibody
CAMTA2 (Calmodulin Binding Transcription Activator 2) is a member of the CAMTA protein family, which regulates transcription by integrating calcium signaling via calmodulin (CaM) interactions . The CAMTA2 antibody is a critical tool for detecting CAMTA2 in research, enabling studies on its roles in cellular processes, disease mechanisms, and transcriptional regulation.
Transcriptional Regulation
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CAMTA2 binds to CGCG motifs in promoters (e.g., Sus2, SBEIc) to activate genes involved in sucrose and starch metabolism in plants .
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In C. elegans and Drosophila, CAMTA homologs regulate neuronal calmodulin (CaM) levels, impacting Ca²⁺ signaling and behavior .
Cancer Biology
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Colon Cancer: CAMTA2 is upregulated and promotes proliferation/metastasis via Wnt/β-catenin signaling. Knockdown reduces tumor growth in vitro and in vivo .
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Tumor Suppression: CAMTA2 interacts with NKX2-2 and GCN5 to regulate stress-responsive miRNAs (e.g., miR-212/132) in pancreatic β-cells, influencing insulin synthesis .
Agricultural Research
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In wheat, CAMTA2 knockout reduces grain size and starch content by downregulating starch synthase (Sus2) and starch-branching enzyme (SBEIc) genes .
Mechanistic Insights
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Protein-Protein Interactions: CAMTA2 recruits histone acetyltransferase GCN5 to enrich H3K9ac/H3K14ac at target gene promoters (e.g., Sus2), enhancing transcriptional activation .
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Subcellular Localization: CAMTA2 localizes to both nucleus and cytoplasm in INS-1 cells, with glucose-dependent interactions with NKX2-2 .
Disease Relevance
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CAMTA2 Dysregulation: High CAMTA2 expression correlates with poor survival in colon cancer patients .
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Neurological Defects: C. elegans camt-1 mutants exhibit Ca²⁺ signaling defects rescued by neuronal CaM supplementation, highlighting CAMTA’s conserved role .
Technical Considerations for CAMTA2 Antibody Use
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Validation: Antibodies are validated using siRNA knockdown (e.g., 50% reduction in Camta1 signal) and peptide-blocking assays .
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Buffers: Store in PBS with 40% glycerol at -20°C; avoid freeze-thaw cycles .
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Controls: Include WT and knockout samples (e.g., camta2 CRISPR mutants) to confirm specificity .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
CAMTA2 is a transcription activator that binds to the DNA consensus sequence 5'-[ACG]CGCG[GTC]-3'. It regulates transcriptional activity in response to calcium signals. CAMTA2 binds calmodulin in a calcium-dependent manner. It plays a role in freezing tolerance in association with CAMTA1 and CAMTA3. In collaboration with CAMTA1 and CAMTA3, CAMTA2 contributes to the positive regulation of the cold-induced expression of DREB1A/CBF3, DREB1B/CBF1 and DREB1C/CBF2. Together with CAMTA3 and CAMTA4, CAMTA2 is involved in the positive regulation of a general stress response. It also participates in tolerance to aluminum. CAMTA2 binds to the promoter of ALMT1 transporter and contributes to the positive regulation of aluminum-induced expression of ALMT1.
- CAMTA2 was identified as an activator of AtALMT1 expression. [CAMTA2] PMID: 25627216
- CAMTA1, CAMTA2 and CAMTA3 collaborate to inhibit salicylic acid biosynthesis at warm temperatures (22 degrees C). These three proteins also work together to positively regulate CBF1, CBF2 and CBF3, ultimately contributing to freezing tolerance. [CAMTA2] PMID: 23581962
Q&A
Experimental Design for CAMTA2 Antibody Validation
Q: How can I validate the specificity and sensitivity of a CAMTA2 antibody in my research? A: To validate the specificity and sensitivity of a CAMTA2 antibody, you should perform several experiments:
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Western Blot: Use the antibody to detect CAMTA2 in cell lysates known to express CAMTA2 and compare with negative controls.
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Immunofluorescence: Perform immunofluorescence on cells or tissues expressing CAMTA2 to verify localization.
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Knockdown/Knockout Experiments: Use siRNA or CRISPR to reduce CAMTA2 expression and verify that the antibody signal decreases accordingly.
Data Interpretation and Contradiction Analysis
Q: How do I handle contradictory results when using CAMTA2 antibodies in different experimental setups? A: Contradictory results can arise from several factors:
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Antibody Specificity: Ensure the antibody is specific to CAMTA2 and not cross-reacting with other proteins.
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Sample Preparation: Verify that sample preparation methods are consistent across experiments.
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Experimental Conditions: Check for differences in experimental conditions such as temperature, pH, or buffer composition.
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Data Analysis: Use statistical methods to assess the significance of observed differences.
Advanced Research Questions: CAMTA2 Function
Q: What role does CAMTA2 play in cellular processes, and how can I investigate its function using CAMTA2 antibodies? A: CAMTA2 is involved in stress responses and transcriptional regulation. To investigate its function:
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Co-immunoprecipitation (Co-IP): Use CAMTA2 antibodies to pull down interacting proteins and identify potential pathways.
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Chromatin Immunoprecipitation (ChIP): Analyze CAMTA2 binding to specific genomic regions to understand its role in gene regulation.
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Cellular Assays: Monitor changes in cellular behavior (e.g., proliferation, differentiation) upon CAMTA2 modulation.
Methodological Considerations for Antibody Conjugation
Q: How can I conjugate CAMTA2 antibodies to other molecules for enhanced detection or therapeutic applications? A: Conjugation of CAMTA2 antibodies to other molecules (e.g., fluorescent dyes, toxins) can be achieved through several methods:
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Chemical Cross-linking: Use reagents like NHS-ester or maleimide to link the antibody to the desired molecule.
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Enzymatic Conjugation: Utilize enzymes like sortase or transglutaminase for site-specific conjugation.
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Genetic Engineering: Express the antibody as a fusion protein with the desired molecule.
Advanced Techniques for CAMTA2 Localization
Q: What advanced techniques can I use to study the subcellular localization of CAMTA2 using CAMTA2 antibodies? A: For detailed localization studies:
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Super-Resolution Microscopy: Techniques like STORM or STED can provide high-resolution images of CAMTA2 distribution.
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Proximity Ligation Assay (PLA): Detect interactions between CAMTA2 and other proteins at the nanoscale.
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Live Cell Imaging: Use fluorescently labeled CAMTA2 antibodies to monitor dynamic changes in localization over time.
CAMTA2 Antibody in Disease Models
Q: How can CAMTA2 antibodies be used in disease models to understand CAMTA2's role in pathogenesis? A: In disease models:
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Immunohistochemistry (IHC): Use CAMTA2 antibodies to assess expression levels in diseased tissues.
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Animal Models: Employ CAMTA2 antibodies to monitor changes in CAMTA2 expression or localization in response to disease progression or treatment.
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Cell Culture Models: Study the effects of CAMTA2 modulation on disease-relevant cellular processes.
Example Data Table: CAMTA2 Antibody Validation
| Method | Sample | Expected Outcome |
|---|---|---|
| Western Blot | Cell Lysate | Specific band at expected molecular weight |
| Immunofluorescence | Fixed Cells | Localization to nucleus or cytoplasm |
| Knockdown/Knockout | siRNA/CRISPR Treated Cells | Reduced signal upon CAMTA2 depletion |
This table illustrates the expected outcomes for validating a CAMTA2 antibody using different methods.
Research Findings: CAMTA2 Function
Recent studies have highlighted the role of CAMTA proteins in stress responses and transcriptional regulation. For instance, CAMTA1 has been implicated in neuronal function and type 2 diabetes, suggesting that CAMTA2 might also play critical roles in similar pathways . Further research using CAMTA2 antibodies can elucidate its specific functions and interactions in these contexts.
