ttc39c Antibody

What is TTC39C and why is it significant for cancer research?

TTC39C is a protein-coding gene located on the long arm of chromosome 18 that contains several putative tetrapeptide repeat (TPR) domains. These domains form structural motifs that promote protein-protein interactions, affecting cell cycle regulation and signaling pathways . TTC39C has gained significance in cancer research because:

• Its expression is significantly upregulated in STK11 mutant lung cancer tissue samples

• It strongly regulates proliferation and metastasis of lung adenocarcinoma cells

• Depletion of TTC39C inhibits proliferation, metastasis, and cloning ability of human lung cancer cells while increasing apoptosis

• It may serve as a potential intervention target for lung cancer treatment

The TPR domains within TTC39C likely mediate interactions between proteins in lung adenocarcinoma patients, making it a valuable target for understanding tumor progression mechanisms.

What are the recommended applications for TTC39C antibodies in research?

Based on validated commercial antibodies, TTC39C antibodies can be reliably used in the following applications:

Researchers should note that optimal dilutions may vary depending on the specific antibody source and experimental conditions. It is always recommended to perform optimization experiments for your specific application.

What species reactivity do commercial TTC39C antibodies have?

Most commercially available TTC39C antibodies show reactivity to human samples . Some antibodies are predicted to cross-react with mouse and rat TTC39C due to sequence homology . When studying TTC39C in non-human models, it's essential to confirm cross-reactivity through preliminary validation experiments before proceeding with full-scale studies.

How can TTC39C antibodies be used to investigate the p53 pathway in lung adenocarcinoma?

TTC39C has been implicated in the p53 pathway in lung adenocarcinoma based on transcriptomic, proteomic, and metabolomic analyses . To investigate this connection:

• Use TTC39C antibodies in co-immunoprecipitation experiments to identify potential binding partners within the p53 pathway

• Perform Western blot analysis using both TTC39C and p53 pathway protein antibodies to assess correlation between expression levels

• Combine TTC39C immunohistochemistry with p53 pathway markers in tumor tissue microarrays to evaluate spatial co-expression patterns

• Design experiments comparing wild-type and p53-deficient cell lines to determine if TTC39C function is p53-dependent

Since the p53 pathway is critical for cell cycle arrest and apoptosis, researchers should design experiments that can distinguish between direct and indirect effects of TTC39C on this pathway.

What experimental considerations are important when studying TTC39C in lung cancer cell models?

When designing experiments to study TTC39C in lung cancer:

• Select appropriate cell lines: A549 and NCI-H1299 have been successfully used in TTC39C research . Note that NCI-H1299 cells are p53-null, which may be relevant when investigating TTC39C's connection to the p53 pathway.

• Gene manipulation approaches:

  • For knockdown studies, shRNA with sequences such as CCGGCGTCTATTGAAGTGTTGTACTCTCGAGAGTACAACACTTCAATAGACGTTTTT have been effective

  • Monitor transfection efficiency using reporter genes (e.g., GFP) and assess after approximately 72 hours

• Functional assays:

  • Proliferation: Celigo and MTT assays can determine effects on cell proliferation

  • Metastasis: Transwell experiments assess metastatic ability

  • Apoptosis: FACS analysis can measure apoptotic effects

  • Clonogenic ability: Clone-formation assays evaluate long-term reproductive viability

• Validation approaches:

  • Confirm knockdown or overexpression at both mRNA level (qPCR) and protein level (Western blot)

  • Use multiple antibody sources or epitopes to ensure consistent results

How do TTC39C expression patterns correlate with patient outcomes in lung adenocarcinoma?

Research using TTC39C antibodies for immunohistochemistry of patient samples has revealed:

• TTC39C expression is significantly increased in lung adenocarcinoma compared to normal lung tissue

• The survival rate of lung adenocarcinoma patients with high TTC39C expression is significantly lower than those with low expression

• Expression patterns may correlate with specific genetic backgrounds, particularly in STK11 mutant cases

When conducting patient outcome studies, researchers should:

• Use validated TTC39C antibodies at appropriate dilutions (1:500-1:1000 for IHC)

• Employ proper quantification methods (e.g., H-scores or percentage of positive cells)

• Account for heterogeneity within tumor samples

• Correlate with clinical data including treatment history and specific genetic alterations

What is the optimal protocol for Western blot analysis using TTC39C antibodies?

For successful Western blot analysis using TTC39C antibodies:

• Sample preparation:

  • Lyse cells or tissues with RIPA buffer

  • Determine protein concentration using BCA protein assay

• Electrophoresis and transfer:

  • Separate proteins using 10-12% SDS-polyacrylamide gels

  • Transfer to PVDF membranes

• Antibody incubation:

  • Block membranes according to antibody manufacturer recommendations

  • Incubate with TTC39C primary antibody at 0.04-0.4 μg/mL or 1:500-1:2000 dilution

  • Incubate overnight at 4°C for optimal results

  • Use appropriate secondary antibody (typically 1:10000 dilution) for 1 hour at room temperature

• Detection and analysis:

  • Visualize using ECL-chemiluminescent kit

  • Quantify band intensities using image analysis software such as Image J

When troubleshooting, the observed molecular weight for TTC39C is approximately 59 kDa .

How should researchers validate TTC39C antibody specificity?

To ensure experimental rigor, validation of TTC39C antibody specificity should include:

• Genetic controls:

  • Compare signal between wild-type cells and TTC39C knockdown/knockout models

  • Use overexpression systems as positive controls

• Peptide competition assays:

  • Pre-incubate antibody with the immunizing peptide or recombinant TTC39C protein

  • Signal should be significantly reduced if the antibody is specific

• Cross-platform validation:

  • Confirm that TTC39C detection by antibody correlates with mRNA expression levels

  • Use multiple antibodies targeting different epitopes of TTC39C

• Epitope information:

  • For some commercial antibodies, the immunogen corresponds to amino acids: TSFHTALELAVDQREIQHVCLYEIGWCSMIELNFKDAFDSFERLKNESRWSQCYYAYLTAVCQGATGDVDGAQIVFKEVQKLFKRKNNQIEQFSV or recombinant fragment corresponding to 62-212 AA of human TTC39C

  • Understanding the epitope region can help predict potential cross-reactivity

What are common challenges when using TTC39C antibodies in immunohistochemistry?

Researchers may encounter these challenges when performing IHC with TTC39C antibodies:

• Fixation and antigen retrieval issues:

  • TTC39C epitopes may be sensitive to fixation conditions

  • Optimize antigen retrieval methods (heat-induced vs. enzymatic)

  • Test different buffer systems for antigen retrieval

• Signal specificity concerns:

  • Use TTC39C-negative tissues as controls

  • Include isotype controls to assess non-specific binding

  • Compare with in situ hybridization for TTC39C mRNA if possible

• Interpretation guidelines:

  • Establish clear scoring criteria for TTC39C positivity

  • Document subcellular localization patterns

  • Consider automated image analysis for quantification

How can researchers integrate TTC39C antibody data with multi-omics approaches?

To maximize insights, researchers can integrate TTC39C antibody-based findings with multi-omics data:

• Correlate protein expression (Western blot/IHC) with:

  • Transcriptomic data: RNA-seq or microarray expression of TTC39C

  • Proteomic data: Mass spectrometry identification of TTC39C and interacting partners

  • Metabolomic data: Altered metabolic pathways associated with TTC39C expression

• Experimental design considerations:

  • Use matched samples across platforms

  • Include appropriate controls for each technique

  • Account for differences in sensitivity between methods

• Data integration approaches:

  • Pathway enrichment analysis incorporating TTC39C interactors

  • Network analysis to identify functional clusters

  • Correlation analysis between TTC39C expression and metabolite levels

This integrated approach can provide insights into how TTC39C affects both energy metabolism and p53 pathways in lung adenocarcinoma .

What are potential applications of TTC39C antibodies in personalized medicine research?

As TTC39C emerges as a potential therapeutic target, antibodies can support personalized medicine research through:

• Patient stratification:

  • Using TTC39C antibodies to identify patients with high vs. low expression

  • Correlating expression patterns with treatment responses

  • Identifying specific patient subgroups (e.g., STK11 mutant cases) that might benefit from TTC39C-targeted therapies

• Companion diagnostic development:

  • Standardizing immunohistochemistry protocols for potential clinical application

  • Determining cutoff values for "high" vs. "low" TTC39C expression

  • Validating across multiple patient cohorts

• Treatment response monitoring:

  • Evaluating changes in TTC39C expression during treatment

  • Correlating with clinical outcomes and resistance mechanisms

How can TTC39C antibodies contribute to understanding protein-protein interactions involving TPR domains?

The TPR domains in TTC39C mediate protein-protein interactions that may be critical for its function in cancer . Researchers can use TTC39C antibodies to:

• Identify interaction partners:

  • Perform co-immunoprecipitation followed by mass spectrometry

  • Conduct proximity ligation assays to visualize interactions in situ

  • Use yeast two-hybrid screening with validation by co-IP

• Characterize domain-specific interactions:

  • Design experiments using truncated TTC39C constructs lacking specific TPR domains

  • Compare interaction profiles between wild-type and mutant TTC39C proteins

  • Assess how post-translational modifications affect TPR domain interactions

• Map interaction interfaces:

  • Use cross-linking approaches coupled with mass spectrometry

  • Employ hydrogen-deuterium exchange mass spectrometry to identify binding regions

  • Validate findings using site-directed mutagenesis

Understanding these interactions may reveal new therapeutic targets within the TTC39C signaling network.