fam110b Antibody

What is FAM110B and why is it significant in cancer research?

FAM110B is a protein belonging to the FAM110 family that has been localized to the centrosome and mitotic spindle. It plays a notable role in cellular processes related to the cell cycle and has demonstrated significant associations with tumor development and progression. Research indicates that FAM110B expression varies considerably across cancer types and holds predictive value for prognosis in several tumors, including brain lower grade glioma (LGG), stomach adenocarcinoma (STAD), and pancreatic adenocarcinoma (PAAD) . Its significance lies in its potential role as a biomarker for immunotherapy response prediction and its involvement in tumor immune microenvironment regulation.

What are the primary applications of FAM110B antibodies in cancer research?

FAM110B antibodies serve multiple crucial research functions:

• Protein Expression Analysis: Antibodies enable detection and quantification of FAM110B across different tumor types through techniques such as western blotting and immunohistochemistry .

• Subcellular Localization Studies: Immunofluorescence staining with FAM110B antibodies allows researchers to investigate its distribution within cellular compartments .

• Prognostic Assessment: Immunohistochemical detection of FAM110B can be utilized to evaluate its correlation with clinical parameters such as TNM staging and lymph node metastasis status .

• Tumor Microenvironment Research: Antibodies facilitate investigation of FAM110B's interactions with immune cell infiltration markers .

What experimental controls should be included when using FAM110B antibodies?

When designing experiments with FAM110B antibodies, incorporate these methodological controls:

• Positive Control: Include samples known to express FAM110B (based on published data, certain NSCLC cell lines like A549, H1299, and LK2 express detectable levels) .

• Negative Control: Utilize samples with confirmed absence or knockdown of FAM110B expression, or use isotype-matched control antibodies to assess non-specific binding.

• Validation Controls: Compare results across multiple detection methods (e.g., RNA-seq data from TCGA compared to protein expression detected via antibodies).

• Specificity Control: Conduct peptide competition assays where the antibody is pre-incubated with the immunizing peptide to confirm binding specificity.

• Loading Controls: For western blots, include housekeeping proteins (β-actin, GAPDH) to normalize expression levels.

Which detection methods are most effective for FAM110B using antibodies?

Different experimental objectives require specific detection approaches:

Research has successfully employed immunohistochemistry to correlate FAM110B expression with clinical outcomes in NSCLC, demonstrating relationships with TNM staging and lymph node metastasis status .

What are the key considerations for optimizing immunohistochemistry protocols with FAM110B antibodies?

For robust IHC results with FAM110B antibodies:

• Antigen Retrieval: Optimize pH and retrieval method (heat or enzymatic) to expose epitopes masked during fixation.

• Antibody Titration: Perform dilution series to determine optimal concentration that maximizes specific signal while minimizing background.

• Incubation Parameters: Test various time/temperature combinations (overnight at 4°C or 1-2 hours at room temperature).

• Detection System Selection: Choose between colorimetric (DAB) or fluorescent detection based on research needs.

• Counterstaining Approach: Adjust hematoxylin concentration for appropriate nuclear contrast without obscuring positive signals.

Previous studies have successfully utilized these approaches to demonstrate that positive FAM110B expression correlates with longer median survival time (56.181±2.348 months compared to 47.701±2.997 months in negative expression cases) .

How should FAM110B antibody validation be performed to ensure specificity?

A comprehensive validation approach includes:

• Cross-reactivity Testing: Evaluate antibody performance across multiple cell lines with varying FAM110B expression levels.

• Knockout/Knockdown Verification: Compare antibody signals in wild-type versus FAM110B knockdown samples using RNAi techniques as demonstrated in NSCLC cell lines .

• Epitope Mapping: Confirm antibody recognition of the intended FAM110B region.

• Orthogonal Method Comparison: Correlate protein detection with mRNA expression data from techniques like RT-qPCR.

• Lot-to-lot Consistency Assessment: Test multiple antibody lots to ensure reproducible performance.

How can FAM110B antibodies be utilized to investigate its role in the tumor microenvironment?

FAM110B's relationship with the tumor microenvironment (TME) can be explored through:

• Multiplex Immunofluorescence: Apply FAM110B antibodies alongside markers for specific immune cell populations (endothelial cells, neutrophils, monocytes/macrophages, and cancer-associated fibroblasts) to evaluate co-localization and spatial relationships .

• Immune Cell Correlation Analysis: Compare FAM110B expression patterns with immune cell infiltration metrics using algorithms such as QUANTISEQ, EPIC, and MCPCOUNTER .

• Single-Cell Analysis Integration: Combine antibody-based detection with single-cell sequencing data to elucidate FAM110B expression across different cell types within the TME .

• Immune Checkpoint Co-expression: Evaluate correlations between FAM110B and immune checkpoint molecules such as PDCD1 and CTLA4 to understand potential implications for immunotherapy .

Research has revealed significant associations between FAM110B expression and infiltration of specific immune cell types, particularly endothelial cells, neutrophils, monocytes/macrophages, and cancer-associated fibroblasts .

What methodological approaches can resolve contradictory data regarding FAM110B expression and function?

When addressing inconsistent experimental results:

• Cell Type Specificity Analysis: FAM110B demonstrates variable expression and function across cancer types. For instance, it shows positive correlations with immune scores in LUAD, ESCA, and COAD but negative correlations in GBM and LGG .

• Context-Dependent Signaling Investigation: Examine FAM110B in relation to specific signaling pathways such as Wnt/β-catenin, which has been implicated in NSCLC .

• Post-translational Modification Assessment: Evaluate potential modifications affecting antibody recognition or protein function.

• Splice Variant Discrimination: Determine if antibodies detect all relevant isoforms or if variant-specific antibodies are needed.

• Microenvironmental Factor Consideration: Assess whether tumor microenvironment factors (hypoxia, pH) affect expression or detection.

How can FAM110B antibodies contribute to understanding its potential as an immunotherapy response biomarker?

To explore FAM110B's biomarker potential:

• Pre/Post-treatment Sample Analysis: Apply antibodies to patient biopsies before and after immunotherapy to track expression changes.

• Correlation with Established Biomarkers: Compare FAM110B expression patterns with tumor mutational burden (TMB) and microsatellite instability (MSI) status .

• Pathway Interaction Mapping: Investigate associations between FAM110B and immunotherapy-relevant pathways through co-immunoprecipitation with pathway components.

• Patient Stratification Method Development: Establish standardized scoring systems for FAM110B expression that correlate with treatment outcomes.

• Liquid Biopsy Application Exploration: Evaluate the feasibility of detecting FAM110B in circulating tumor cells as a non-invasive monitoring approach.

Research has identified significant correlations between FAM110B expression and TMB/MSI status across multiple cancer types, with notable negative correlations in BRCA, ESCA, STAD, STES, HNSC, and THCA .

What experimental approaches can determine FAM110B's mechanistic role in cancer progression?

To elucidate FAM110B's functional significance:

• Gene Modulation Studies:

  • Overexpression models in cancer cell lines have demonstrated that FAM110B can inhibit proliferation and invasion in NSCLC cell lines (A549, H1299, LK2) .

  • RNAi knockdown approaches can reveal loss-of-function effects .

• Pathway Analysis:

  • Western blot analysis of Wnt/β-catenin pathway components following FAM110B modulation can identify downstream effectors .

  • Antibodies can be used to confirm pathway alterations at the protein level.

• In Vivo Models:

  • Xenograft models with FAM110B-modulated cell lines allow for assessment of tumor growth and metastatic potential.

  • Immunohistochemical analysis of resulting tumors using FAM110B antibodies can confirm expression maintenance.

• Drug Sensitivity Correlation:

  • Combine FAM110B expression analysis with drug screening to identify associations with specific inhibitors (e.g., γ-Secretase Inhibitor I, AICAR, GW843682X from GDSC dataset) .

How can FAM110B antibodies be integrated into multi-omics cancer research approaches?

For comprehensive multi-omics integration:

• Proteogenomic Correlation:

  • Compare protein-level detection via antibodies with genomic and transcriptomic data from platforms like TCGA and GTEx .

  • Identify discrepancies between mRNA and protein levels that suggest post-transcriptional regulation.

• Epigenetic-Proteomic Analysis:

  • Correlate FAM110B protein expression with methylation patterns from platforms like GSCALite .

  • Investigate how epigenetic modifications affect antibody-detectable protein levels.

• Spatial Transcriptomics-Immunohistochemistry Alignment:

  • Combine spatial transcriptomic data with immunohistochemical detection to map regional expression variations.

  • Validate transcript-level findings with protein-level confirmation.

• Single-Cell Multi-Modal Analysis:

  • Integrate single-cell RNA-seq with protein detection using FAM110B antibodies to correlate transcription with translation at individual cell resolution .

What methodological considerations apply when studying FAM110B in different cancer types?

Cancer-specific approach adjustment is essential:

• Expression Baseline Establishment:

  • FAM110B expression varies significantly across cancer types, requiring appropriate control selection .

  • For example, in NSCLC research, compare expression in tumor tissue with paired normal lung tissue .

• Prognostic Relevance Assessment:

  • Adapt analytical methods based on cancer-specific prognostic patterns:

  • In NSCLC, FAM110B positivity correlates with better outcomes (median survival 56.181±2.348 vs 47.701±2.997 months) .

  • In other cancers, different survival correlations may exist.

• Microenvironment Context Consideration:

  • Evaluate cancer-specific immune infiltration patterns in relation to FAM110B.

  • For example, COAD, READ, PAAD, and UVM show positive correlations with markers of M1/M2 macrophages, endothelial cells and CAFs, while LGG shows opposite patterns .

• Antibody Validation Requirements:

  • Confirm antibody performance in the specific cancer tissue type being studied.

  • Consider fixation and processing variables that may affect epitope recognition.

What are common technical issues when using FAM110B antibodies and their solutions?

How can researchers address antibody cross-reactivity with other FAM110 family members?

To ensure FAM110B-specific detection:

• Epitope Selection Verification: Confirm the antibody targets unique regions not conserved among FAM110A, FAM110C, and FAM110D.

• Recombinant Protein Controls: Test antibody against all recombinant FAM110 family proteins to assess cross-reactivity.

• Sibling Gene Knockdown: Perform selective knockdown of each family member to confirm antibody specificity.

• Antibody Absorption Testing: Pre-absorb antibodies with recombinant FAM110 family proteins to reduce cross-reactivity.

• Western Blot Molecular Weight Verification: Confirm detection at FAM110B-specific molecular weight (approximately 46 kDa) versus other family members.

What considerations apply when selecting FAM110B antibodies for different experimental applications?

Application-specific selection criteria include:

• Antibody Format Considerations:

  • Monoclonal antibodies offer consistent reproducibility for quantitative applications

  • Polyclonal antibodies may provide stronger signals for detection of low-abundance proteins

• Epitope Accessibility Assessment:

  • For IHC/IF: Select antibodies targeting epitopes that remain accessible after fixation

  • For IP: Choose antibodies recognizing native conformations

  • For WB: Prioritize antibodies detecting denatured epitopes

• Species Cross-Reactivity Requirements:

  • For translational research: Confirm reactivity across relevant species (human, mouse, etc.)

  • For clinical applications: Human-specific antibodies may be preferred

• Clone Selection Strategy:

  • Validate multiple clones for consistent results

  • Consider clones recognizing different epitopes for confirmation of specificity