RUB3 Antibody
Product Specs
Target Background
Q&A
Here’s a structured collection of FAQs tailored for researchers studying RUNX3 Antibody (note: corrected from "RUB3" based on scientific literature), incorporating experimental design, methodological insights, and data analysis challenges:
What is the biological role of RUNX3, and how do antibodies targeting it aid in mechanistic studies?
RUNX3 is a transcription factor critical for immune cell differentiation (e.g., CD8+ T cells, NK cells) and tumor suppression . Antibodies like R3-5G4 enable:
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Flow cytometry: Tracking RUNX3 expression in immune subsets (e.g., intraepithelial lymphocytes) .
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Immunohistochemistry: Localizing RUNX3 in gastric or neural tissues to study tumorigenesis .
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Knockdown validation: Confirming CRISPR/Cas9-mediated RUNX3 deletion via Western blot .
What validation criteria ensure RUNX3 antibody specificity in experimental models?
Key validation steps include:
How do researchers optimize RUNX3 antibody dilution for diverse applications?
A tiered approach is recommended:
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Pilot titration: Test dilutions (1:100–1:1,000) in control tissues.
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Blocking validation: Use peptide competition to confirm specificity .
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Cross-species reactivity: Validate in murine/human models (e.g., thymic sections) .
How can epitope mapping resolve contradictory RUNX3 localization data across studies?
Discrepancies often arise from non-overlapping epitopes. Strategies include:
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Structural alignment: Compare antibody-binding regions to RUNX3 domains (e.g., Runt homology domain) .
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Mutagenesis: Introduce point mutations (e.g., D127A) to disrupt epitope binding .
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Multiplex assays: Combine IF, ChIP-seq, and Co-IP to reconcile nuclear vs. cytoplasmic signals .
What computational approaches improve RUNX3 antibody design for cross-reactive epitopes?
Lessons from SARS-CoV-2 antibody engineering :
How do researchers address RUNX3 antibody cross-reactivity in autoimmune disease models?
Systemic sclerosis (SSc) studies reveal:
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Interference mitigation: Pre-adsorb serum with RUNX1/RUNX2 to isolate RUNX3-specific signals .
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Assay thresholds: Establish cutoff values (e.g., >40 Units = moderate positivity) .
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Longitudinal monitoring: Track antibody titers in SSc patients with renal crisis .
Why do RUNX3 expression levels vary between flow cytometry and Western blot data?
Common pitfalls and solutions:
