RNR1 Antibody

Here’s a structured collection of FAQs for academic researchers studying RNR1 antibodies, synthesized from peer-reviewed literature and patent databases:

Advanced Research Questions

How should researchers resolve contradictions in RNR1 localization studies between cytoplasmic and nuclear compartments?

• Analytical framework:

  • Temporal analysis: Collect time-course data post-DNA damage. RNR1 relocalizes to nuclei during S-phase arrest .

  • Fractionation controls: Validate subcellular isolation purity using compartment-specific markers (e.g., Lamin B1 for nuclei).

  • Antibody validation: Compare multiple clones (e.g., N-terminal vs. C-terminal epitopes) to rule out epitope masking .

What orthogonal approaches complement RNR1 antibody-based assays when studying dNTP pool regulation?

• Integrated workflow:

  • Metabolomics: Quantify dATP/dTTP via LC-MS alongside RNR1 WB.

  • Genetic interaction mapping: Combine RNR1 overexpression with dun1Δ mutants to assess checkpoint pathway dependencies .

  • Single-cell imaging: Use Fucci cell cycle reporters to correlate RNR1 levels with S-phase progression.

How can researchers design studies to assess RNR1's role in chemotherapy resistance while controlling for off-target immune effects?

• Experimental design:

  • Use syngeneic tumor models with RNR1 conditional knockouts.

  • Pair anti-RNR1 therapies with PD-1 axis antagonists (e.g., cemiplimab) to decouple direct metabolic effects from immune checkpoint modulation .

  • Monitor dNTP salvage pathways via TK1/DCK expression arrays to identify compensatory mechanisms.

Methodological Notes

• Antibody validation: For studies using RNR1 inhibitors (e.g., triapine), pre-treat lysates with 2 mM DTT to reduce disulfide-linked aggregates that cause false-positive WB bands .

• Multiplex assays: When combining RNR1 IF with EdU labeling, fix cells with methanol (not paraformaldehyde) to preserve both epitope accessibility and DNA incorporation signals .