RAD52-1 Antibody
Description
RAD52 Antibody Overview
RAD52 antibodies target the human RAD52 protein (UniProt: P43351), which facilitates DNA strand annealing, single-strand annealing (SSA), and HR-mediated repair . These antibodies are used to investigate RAD52's nuclear foci formation, interaction with DNA repair complexes, and therapeutic targeting in BRCA-deficient cancers .
Key RAD52 Antibodies and Their Applications
Below is a comparative analysis of widely used RAD52 antibodies:
Role in BRCA-Deficient Cells
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RAD52 forms nuclear foci co-localizing with γH2AX in BRCA1/2-deficient cells, as shown using HA-tagged RAD52 1–209 constructs .
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Depletion of RAD52 sensitizes BRCA2-deficient cells to G-quadruplex (G4)-stabilizing compounds (e.g., pyridostatin), leading to DSB accumulation .
Mechanistic Insights
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Mediator Function: RAD52 facilitates XPF recruitment to process DNA secondary structures at DSB ends, enabling HR repair .
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Synthetic Lethality: Inhibitors like C791-0064 disrupt RAD52 self-association, selectively killing BRCA2-deficient cells (IC50: 28.92 μM vs. 64.62 μM in BRCA2-proficient cells) .
Technical Considerations for RAD52 Antibodies
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Western Blotting: Most antibodies detect RAD52 at ~46–55 kDa, with variations due to post-translational modifications (e.g., SUMOylation) .
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Immunofluorescence: Optimal results require PFA/Triton X-100 fixation, as validated in U2OS and MDA-MB-436 cells .
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Knockdown Validation: Antibodies like NBP2-58116 and 28045-1-AP are validated in CRISPR-Cas9 KO models .
Limitations and Future Directions
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research has demonstrated the role of the ODB1 protein in group-II intron splicing within the mitochondria of Arabidopsis. PMID: 26048959
- Arabidopsis odb1 mutants, while indistinguishable from wild-type in morphology, exhibited reduced mitochondrial homologous recombination activity following DNA damage induced by genotoxic stress. PMID: 22762281
Q&A
FAQs for RAD52-1 Antibody in Academic Research
Advanced Research Questions
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How can RAD52-1 antibody resolve contradictions in RAD52’s mediator role in RAD51 nucleoprotein filament (NPF) formation?
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Experimental design:
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Data interpretation:
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What methods leverage RAD52-1 antibody to study phase-separated repair condensates?
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Proximity ligation assay (PLA): Detect RAD52-RAD51 interactions in condensates using Duolink® reagents .
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Correlative light-electron microscopy (CLEM): Combine antibody-based fluorescence with EM to visualize condensate ultrastructure .
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FRAP analysis: Measure recovery kinetics of GFP-tagged RAD52 after photobleaching in live cells .
Key findings:
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Methodological Challenges & Solutions
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How to troubleshoot non-specific bands in RAD52-1 Western blots?
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Can RAD52-1 antibody distinguish between monomeric vs. oligomeric RAD52 states?
Data Integration Table
Comparative roles of RAD52 across studies:
Critical Analysis
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Contradictions in mediator activity: While reports RAD52 supports RAD51 HR in BRCA2-/- cells, shows condensate-driven RAD51 clustering is BRCA2-independent. Resolve by testing RAD52-1 localization in isogenic BRCA2 knockout lines with/without RAD52 inhibitors .
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Antibody limitations: Commercial RAD52-1 antibodies often fail to detect phosphorylation at Serine 14 (pS14-RAD52), critical for condensate formation. Combine with Phos-tag® gels for post-translational modification analysis .
