STK32C Antibody

What is STK32C and why is it important in research?

STK32C (Serine/Threonine Kinase 32C), also known as PKE or YANK3, belongs to the serine/threonine protein kinase AGC superfamily. It consists of 486 amino acids and contains a protein kinase domain . STK32C uses magnesium as a cofactor to catalyze the conversion of ATP to ADP while transferring phosphate groups to target proteins . This protein was initially found to be highly expressed in brain tissues but has gained significant research interest due to its involvement in various cancer types, notably bladder cancer . The phosphorylation and dephosphorylation of serine and threonine residues by such kinases regulate fundamental cellular functions including cell division, homeostasis, and apoptosis, making STK32C a critical target for investigation .

What types of STK32C antibodies are available for research?

Several types of STK32C antibodies are available for research purposes, each with specific characteristics and applications:

• Mouse monoclonal antibodies: The STK32C Antibody (18-56) is an IgG2a κ mouse monoclonal antibody that can detect mouse, rat, and human STK32C .

• Rabbit polyclonal antibodies: Products like ab153917 are rabbit polyclonal antibodies raised against recombinant fragment protein within human STK32C aa 100-450 .

These antibodies are provided in various formats, including non-conjugated forms, allowing researchers flexibility in experimental design based on their specific requirements .

What detection methods are compatible with STK32C antibodies?

STK32C antibodies are compatible with multiple detection methods, enabling versatile research applications:

Different antibody products may have specific optimal dilutions, so researchers should refer to product-specific documentation for best results . The diversity of compatible methods allows researchers to investigate STK32C expression, localization, and interactions across various experimental contexts.

How should immunohistochemistry be optimized for STK32C detection in tissue samples?

For optimal immunohistochemical detection of STK32C in tissue samples, researchers should follow these methodological considerations:

• Sample preparation: Tissues should be fixed in 10% neutral-buffered formalin and embedded in paraffin. Sections of approximately 4-μm thickness are recommended for consistent results .

• Antigen retrieval: Use citrate buffer (pH = 6.0) combined with 1.5% hydrogen peroxide in methanol to unmask antigens that may have been cross-linked during fixation .

• Primary antibody incubation: Incubate slides with anti-STK32C antibody (typically at 1:200 dilution for rabbit antibodies) overnight at 4°C for optimal binding .

• Secondary antibody and visualization: Apply the appropriate secondary antibody for 1 hour at room temperature, followed by 3,3′-diaminobenzidine for visualization of the signal .

• Scoring and analysis: For semi-quantitative assessment, the H-score approach is recommended, which considers both staining intensity and the proportion of positively stained cells . The formula Z = A × B is used, where A represents the proportion score (0-4) and B represents the intensity score (0-3) .

This methodology has been validated in studies examining STK32C expression in bladder cancer tissues, where researchers successfully identified correlations between expression levels and clinical features .

What are the best practices for knockdown studies targeting STK32C?

When designing knockdown experiments to study STK32C function:

• Vector selection: STK32C-RNAi lentiviral vectors have proven effective in previous studies. Control lentiviral vectors (shCtrl) should be used in parallel to establish baseline measurements .

• Verification of knockdown: Always verify the efficacy of knockdown using both real-time quantitative PCR and Western blot to confirm reduction at both mRNA and protein levels .

• Cell selection: Following transfection, qualified cells should be selected using appropriate selection markers before proceeding with functional assays .

• Off-target effect control: To exclude potential off-target effects that might confound your results, include self-rescue experiments where STK32C expression is restored in knockdown cells .

• Functional assays: Compare biological behaviors between shSTK32C cells and shCtrl cells, focusing on phenotypes relevant to your research question (e.g., proliferation, migration, invasion for cancer studies) .

Studies employing these methodologies have successfully demonstrated that silencing STK32C inhibits tumor cell proliferation, migration, and invasion in vitro, and restricts tumor growth in mouse models .

How is STK32C expression altered in cancer, and what are the implications?

Analysis of TCGA (The Cancer Genome Atlas) data has revealed significant alterations in STK32C expression across multiple cancer types:

• Bladder cancer: STK32C is significantly overexpressed in bladder tumors compared to adjacent normal tissues (fold change = 2.531; P < 0.001) . In paired sample analyses, 12 out of 19 samples showed up-regulation of STK32C .

• Other cancer types: STK32C is also overexpressed in breast invasive carcinoma (BRCA), esophageal carcinoma (ESCA), head and neck squamous cell carcinoma (HNSC), liver hepatocellular carcinoma (LIHC), lung adenocarcinoma (LUAD), stomach adenocarcinoma (STAD), and prostate adenocarcinoma (PRAD) . Interestingly, it is downregulated in lung squamous cell carcinoma (LUSC) .

• Genetic alterations: Mutations and amplifications of the STK32C gene are rare, with only R97W mutation documented in some TCGA patients. Amplification of the STK32C gene did not correlate with expression levels .

The overexpression of STK32C across multiple cancer types suggests it functions as an active contributor to oncogenesis and tumor progression, making it a potential therapeutic target and prognostic marker .

What is the relationship between STK32C expression and clinical outcomes in cancer patients?

Immunohistochemical studies of bladder cancer patients have established significant correlations between STK32C expression and clinical outcomes:

• Clinical associations: High STK32C expression is significantly associated with clinicopathological characteristics including older age, male gender, larger tumor size, multiple tumors, advanced pTNM stage, and higher recurrence rates .

• Subcellular localization: Both nuclear and cytoplasmic localization of STK32C has been observed in tumor cells, with higher pathologic T-stage bladder cancers showing more pronounced nuclear staining . This suggests potential stage-dependent nuclear translocation that may influence disease progression.

• Survival impact: Kaplan-Meier survival analyses demonstrate that high levels of STK32C are significantly associated with poor recurrence-free survival (RFS) in patients treated with transurethral resection of bladder tumor (TURBT) .

These findings suggest that STK32C expression assessment could serve as a valuable prognostic biomarker in bladder cancer management, potentially guiding treatment decisions and follow-up protocols .

What molecular pathways does STK32C interact with in cancer progression?

Research has identified several key pathways and mechanisms through which STK32C contributes to cancer progression:

• HMGB1 pathway: Microarray analysis revealed that silencing STK32C inhibits the activity of the HMGB1 (High Mobility Group Box 1) pathway in bladder cancer . STK32C regulates the expression of key genes within this pathway, which is known to promote inflammation and cancer progression.

• Immune regulation: Data mining from different studies has shown that STK32C gene expression changes correlate with CD4+ regulatory T-cell function and lung inflammation , suggesting potential immunomodulatory roles.

• Cell proliferation and invasion: Functional studies have demonstrated that knockdown of STK32C inhibits tumor cell proliferation, migration, and invasion in vitro . This suggests STK32C influences pathways controlling these hallmark cancer processes.

Understanding these molecular interactions provides opportunities for developing targeted therapeutic strategies that disrupt STK32C-mediated oncogenic signaling .

How should researchers quantify and analyze STK32C expression in experimental settings?

For rigorous quantification and analysis of STK32C expression:

• In tissue samples: The H-score approach is recommended for semi-quantitative assessment of immunohistochemical staining. This system accounts for both staining intensity (0-3) and the proportion of positively stained cells (0-4). The final score Z is calculated by multiplying these values (Z = A × B) .

• Classification thresholds: Based on the distribution of final scores, samples can be categorized into high expression (Z > 3) and low expression (Z ≤ 3) groups for statistical analysis and correlation with clinical parameters .

• Statistical analysis: SPSS or similar statistical software should be used for data analysis. Quantitative data should be assessed by mean ± SD. Choose parametric or non-parametric testing based on whether data shows variance heterogeneity .

• Associations analysis: Use chi-square tests to analyze associations between STK32C expression and clinicopathologic features. For survival analyses, apply Kaplan-Meier methods to evaluate relationships between STK32C expression and outcomes like recurrence-free survival .

• Significance threshold: A P-value < 0.05 is generally considered statistically significant for differences between groups .

This systematic approach to quantification enables reliable comparison across studies and facilitates the identification of clinically relevant associations .

What are the experimental challenges in studying STK32C function?

Researchers investigating STK32C function face several experimental challenges:

• Isoform complexity: STK32C exists in two isoforms due to alternative splicing events . Experiments must account for potential isoform-specific functions and expression patterns.

• Subcellular localization variability: STK32C exhibits both nuclear and cytoplasmic localization, with patterns varying by cancer stage . This requires careful selection of experimental methods that can distinguish between compartment-specific roles.

• Tissue specificity: While originally identified as highly expressed in brain tissues, STK32C shows variable expression across different tissue types and cancer subtypes . This necessitates tissue-specific optimization of detection protocols.

• Functional redundancy: As a member of the serine/threonine protein kinase family, STK32C may share functional redundancy with related kinases, complicating the interpretation of knockdown phenotypes.

• Target identification: Identifying the specific protein targets of STK32C phosphorylation activity remains challenging but is crucial for understanding its mechanistic role in cancer progression.

Addressing these challenges requires careful experimental design, appropriate controls, and the integration of multiple methodological approaches to build a comprehensive understanding of STK32C biology.

What are promising avenues for future STK32C research in cancer therapeutics?

Based on current knowledge, several promising research directions emerge for STK32C-focused cancer therapeutics:

• Inhibitor development: Given STK32C's role in promoting tumor progression, developing specific small molecule inhibitors targeting its kinase activity could yield new therapeutic options for cancers with STK32C overexpression .

• Combination therapies: Investigating the potential synergistic effects of combining STK32C inhibition with existing chemotherapeutic regimens could enhance treatment efficacy for aggressive cancers.

• Biomarker validation: Further validation of STK32C as a prognostic biomarker across larger patient cohorts and diverse cancer types would strengthen its clinical utility for risk stratification and treatment planning .

• Pathway interactions: Deeper investigation of the relationship between STK32C and the HMGB1 pathway could reveal additional therapeutic targets and clarify the broader impact of STK32C inhibition on inflammatory and immune responses in the tumor microenvironment .

• Resistance mechanisms: Studying potential mechanisms of resistance to STK32C-targeted therapies would be valuable for developing effective sequential or combinatorial treatment strategies.

The significant associations between STK32C expression and poor clinical outcomes in bladder cancer patients suggest that developing therapeutic approaches targeting this kinase could potentially improve patient survival and quality of life .