BRCA2A Antibody
Description
BRCA2 Protein Overview
The BRCA2 protein (384 kDa) is encoded by the BRCA2 gene located on chromosome 13q12.3. Key functions include:
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DNA Repair: Facilitates homologous recombination (HR) by recruiting RAD51 to single-stranded DNA (ssDNA), enabling accurate repair of double-strand breaks (DSBs) .
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Tumor Suppression: Loss-of-function mutations increase risks for breast, ovarian, pancreatic, and prostate cancers .
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Cytokinesis Regulation: May assist in cell division processes, though this role is less characterized .
Key BRCA2 Antibodies and Their Applications
Below is a comparative analysis of widely used BRCA2 antibodies, validated across diverse experimental systems:
DNA Repair Mechanisms
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BRCA2 deficiency impairs HR, leading to error-prone repair via nonhomologous end joining (NHEJ) .
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Key Evidence: BRCA2 reconstitution in Capan-1 cells increased HR efficiency by 10-fold, restoring RAD51-mediated DSB repair .
Therapeutic Implications
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PARP Inhibitors: BRCA2-deficient cells show hypersensitivity to PARP inhibitors (e.g., olaparib) due to synthetic lethality .
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Immunotherapy: BRCA2-deficient tumors exhibit enhanced response to immune checkpoint blockade (ICB) compared to BRCA1-deficient tumors, attributed to elevated immune infiltration .
Experimental Models
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5C6 Antibody Study: The lupus autoantibody 5C6 induced DNA damage (γH2AX foci) selectively in BRCA2-deficient DLD1 cells, reducing proliferation by 41% .
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MANO-B Assay: High-throughput screening of 244 BRCA2 variants identified pathogenic mutations with 95% sensitivity/specificity using PARP inhibitor sensitivity as a readout .
Technical Considerations
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Western Blot: BRCA2 migrates at ~380 kDa; reducing conditions are critical to prevent aggregation .
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IHC Optimization: Heat-induced epitope retrieval (HIER) with citrate buffer improves staining in FFPE tissues .
Future Directions
Product Specs
Target Background
- AtBRCA2 is crucial for proper meiotic synapsis and facilitates the recruitment of AtRAD51 and AtDMC1. (PMID: 22077663)
- Evidence indicates that the BRCA2-RAD51 complex, known for its role in homologous recombination (HR), also directly participates in transcriptional regulation during plant immune responses. (PMID: 21149701)
- Inefficient DNA repair in the *atbrca2a-/atbrca2b* double mutant disrupts the cell cycle in apical meristems. (PMID: 19457980)
- Brca2 plays an essential role in Arabidopsis meiosis. (PMID: 15014444)
Q&A
Experimental Design for BRCA2 Antibody Validation
Question: What methods should be used to validate the specificity and sensitivity of BRCA2 antibodies in experimental settings?
Answer: Validation of BRCA2 antibodies involves several key methods:
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Western Blot (WB): Use WB to assess antibody specificity by detecting BRCA2 protein in cell lysates.
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Immunohistochemistry (IHC): Perform IHC on tissue samples to evaluate antibody performance in fixed tissues.
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Immunoprecipitation (IP) and Mass Spectrometry (MS): Conduct IP-MS to confirm antibody specificity by identifying co-precipitated proteins.
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Chromatin Immunoprecipitation (ChIP): Use ChIP to study BRCA2's role in chromatin remodeling and DNA repair.
Data Contradiction Analysis in BRCA2 Research
Question: How can researchers address data inconsistencies when using different BRCA2 antibodies across various studies?
Answer: Addressing data inconsistencies involves:
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Cross-validation: Use multiple antibodies across different experimental platforms to confirm findings.
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Literature Review: Compare results from similar studies to identify patterns or discrepancies.
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Methodological Comparison: Analyze differences in experimental conditions, such as cell lines or tissue types, that might contribute to inconsistencies.
Advanced Research Questions: BRCA2 Function in DNA Repair
Question: What role does BRCA2 play in homologous recombination, and how can this be studied using BRCA2 antibodies?
Answer: BRCA2 is crucial for homologous recombination (HR), a process essential for DNA repair. To study this:
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Immunofluorescence (IF): Use IF to visualize BRCA2 localization at DNA damage sites.
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Cellular Assays: Perform assays that measure HR efficiency, such as DR-GFP or SSA assays, in cells with manipulated BRCA2 expression.
Methodological Considerations for BRCA2 Antibody Selection
Question: What criteria should researchers use when selecting a BRCA2 antibody for their studies?
Answer: When selecting a BRCA2 antibody, consider:
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Specificity: Ensure the antibody specifically targets BRCA2 without cross-reacting with other proteins.
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Sensitivity: Choose an antibody that can detect BRCA2 in the desired experimental conditions.
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Application Suitability: Select antibodies validated for the intended application (e.g., WB, IHC, IF).
Data Interpretation Challenges in BRCA2 Research
Question: How can researchers interpret complex data from BRCA2 antibody experiments, especially when dealing with conflicting results?
Answer: Interpreting complex data involves:
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Statistical Analysis: Use appropriate statistical methods to analyze data and identify significant trends.
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Control Experiments: Include robust controls to validate experimental conditions and antibody performance.
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Literature Contextualization: Place findings within the broader context of existing BRCA2 research to understand potential implications.
Advanced Techniques for BRCA2 Antibody-Based Research
Question: What advanced techniques can be employed to further elucidate BRCA2's role in cancer using BRCA2 antibodies?
Answer: Advanced techniques include:
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Single-Molecule Localization Microscopy (SMLM): Use SMLM to study BRCA2's spatial organization at the nanoscale.
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Proximity Ligation Assay (PLA): Perform PLA to detect protein-protein interactions involving BRCA2.
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CRISPR-Cas9 Gene Editing: Use CRISPR-Cas9 to manipulate BRCA2 expression and study its effects on cellular processes.
Example Data Table: BRCA2 Antibody Validation
| Antibody ID | Application | Specificity | Sensitivity |
|---|---|---|---|
| Ab216972 | WB, IHC | High | Moderate |
| Ab123456 | IF, ChIP | Moderate | High |
This table illustrates how different antibodies perform across various applications, helping researchers choose the most suitable antibody for their studies.
