ORRM6 Antibody

Based on analysis of available research materials, the following FAQs address key considerations for working with ORC6 antibodies in academic research contexts. (Note: The query mentions "ORRM6," but available literature focuses on ORC6 [Origin Recognition Complex subunit 6], a validated target in oncology and immunology research.)

What experimental controls are essential when using ORC6 antibodies in immunofluorescence (IF)?

Include these controls to ensure data reliability:

• Positive controls: Use cell lines with confirmed ORC6 expression (e.g., hepatocellular carcinoma [LIHC] or glioma lines) . Validate antibody specificity via CRISPR/Cas9 KO models .

• Negative controls:

  • Secondary antibody-only samples to detect non-specific binding3.

  • PBS-only treatment to assess autofluorescence (common in stressed or necrotic tissues)3.

• Endogenous controls: Monitor DNA damage markers (e.g., γH2AX) or mitochondrial morphology to rule out experimental artifacts3.

How does ORC6 influence baseline cellular processes relevant to cancer?

ORC6 is critical for DNA replication and genome stability. Key findings include:

How to resolve contradictions in ORC6’s role across tumor types?

ORC6 exhibits tissue-specific effects:

• Pro-tumorigenic in LIHC/glioma: Correlates with immunosuppression (reduced Tem CD8+ cells) and poor prognosis .

• Ambiguous in THCA/KIRC: Associates with immune cell infiltration but no clear survival link .
  Methodological recommendations:

  • Use multi-cohort validation (e.g., CGGA, ICGC-LIRI-JP) to contextualize findings .

  • Perform pathway enrichment (GSEA) to identify tumor-specific signaling networks .

What mechanisms link ORC6 to immunotherapy resistance?

ORC6 modulates immune checkpoint interactions:

• Immune checkpoint correlation:

  Checkpoint	Correlation with ORC6	Therapeutic Implication
  PD-L1	Positive in glioma	May limit PD-1 inhibitor efficacy
  CTLA-4	Negative in LIHC	Potential synergy with CTLA-4 blockade

• TIDE analysis: ORC6 predicts resistance to ICB therapy in 7 cohorts (melanoma, bladder cancer) .

How to validate ORC6’s role in NFκB-driven inflammation?

Stepwise approach:

• In vitro models: Use THP-1 macrophages with ORC6 KO/overexpression + LPS stimulation .

• Key readouts:

  • p65 nuclear translocation (IF/confocal microscopy) .

  • IL-6/TNF-α ELISA (serum/tissue lysates) .

• Pharmacological inhibition: Combine ORC6 silencing with BMS-345,541 (NFκB inhibitor) to confirm pathway dependence .

Methodological Notes

• Antibody validation: Always cross-verify using orthogonal methods (e.g., Western blot with KO controls , spatial transcriptomics in FFPE tissues).

• Data integration: Leverage multi-omics databases (TISMO, TCGA) to correlate ORC6 expression with immune metrics .