AURKB Antibody

What is Aurora Kinase B and what are its primary cellular functions?

Aurora Kinase B (AURKB) is a 39 kDa serine/threonine protein kinase that functions as a critical component of the chromosomal passenger complex (CPC). This complex acts as a key regulator of mitosis and is essential for accurate chromosome segregation, cytokinesis, protein localization to the centromere and kinetochore, correct microtubule-kinetochore attachment, and regulation of the mitotic checkpoint .

AURKB forms a tight complex with inner centromere protein and survivin. Research demonstrates that inactivation of any of these proteins results in similar defects in chromosome segregation, highlighting their interdependent roles in maintaining genomic stability during cell division .

How does AURKB contribute to cancer development and progression?

AURKB has been revealed through various studies to play a significant role in cancer development and progression, though the specific mechanisms remain incompletely understood . Significant overexpression of AURKB has been documented in numerous human tumors including non-small cell lung carcinoma, astrocytoma, seminoma, and carcinomas of the colon, prostate, endometrium, and thyroid .

The expression level of AURKB is associated with cell proliferation and prognosis in these tumors, suggesting its potential role in promoting aberrant cell division and genomic instability that contribute to malignant transformation and tumor progression .

What are the optimal protocols for immunohistochemical detection of AURKB in tissue samples?

Based on validated research protocols, optimal immunohistochemical detection of AURKB includes:

• Tissue preparation: Bake paraffin sections at 65°C for 1 hour

• Deparaffinization: Sequential xylene dewaxing and gradient ethanol dehydration

• Antigen retrieval: High-temperature antigen retrieval method

• Blocking: Incubate with 3% hydrogen peroxide for 10 minutes

• Primary antibody: Apply diluted AURKB antibody (1:200 dilution) and incubate for 24 hours

• Secondary antibody: Incubate with working solution of secondary antibody for 20 minutes at room temperature

• Detection: Perform DAB color development and hematoxylin re-staining

• Mounting: Dehydrate and seal slides

For staining interpretation, researchers should evaluate both intensity and proportion of stained cells using the following criteria:

• Staining intensity: 0 (no staining), 1 (light yellow), 2 (dark yellow), 3 (yellow-brown)

• Proportion of stained cells: 0 (0%), 1 (1-25%), 2 (26-50%), 3 (51-75%), 4 (76-100%)

• Positivity determination: <2 points is negative; ≥2 points is positive

What factors should be considered when selecting fluorescent conjugates for AURKB antibodies?

When selecting fluorescent conjugates for AURKB antibodies, researchers should consider:

• Target abundance: For low-abundance targets, avoid blue fluorescent dyes like CF®405S and CF®405M as they have lower fluorescence intensity and can give higher non-specific background than other dye colors .

• Spectral properties: Choose dyes that are compatible with your imaging system and other fluorophores in multiplex experiments.

• Photostability: Consider the photostability of fluorescent conjugates, particularly for extended imaging sessions. CF® dyes, for example, offer exceptional brightness and photostability .

• Application compatibility: Ensure the conjugated antibody has been validated for your specific application (immunofluorescence, flow cytometry, etc.).

• Lead time considerations: Because suppliers offer numerous antibody and conjugation options, primary antibody conjugates may be made to order with varying lead times (typically up to one week for CF® dye and biotin conjugates, and up to 2-3 weeks for fluorescent protein and enzyme conjugates) .

How can researchers validate the specificity of AURKB antibodies in their experimental systems?

To ensure antibody specificity for AURKB detection:

• Western blot validation: Confirm a single band of approximately 39 kDa, which corresponds to the molecular weight of AURKB .

• Knockout/knockdown controls: Compare staining patterns in AURKB-depleted samples (via siRNA, CRISPR, etc.) with control samples to verify specificity.

• Tissue controls: Include positive controls (tissues known to express high levels of AURKB, such as certain cancer tissues) and negative controls (tissues with minimal AURKB expression) .

• Antibody validation protocols: Implement comprehensive validation protocols including peptide competition assays, where pre-incubation of the antibody with the immunizing peptide should abolish specific staining.

• Multiple antibody approach: Use antibodies from different clones targeting distinct epitopes to confirm consistent staining patterns.

What patterns of AURKB expression have been observed across different cancer types?

Analysis of AURKB expression across cancer types reveals consistent overexpression in multiple malignancies. According to TIMER 2.0 database analysis, AURKB mRNA is significantly overexpressed in 18 cancer types compared to normal tissues, including:

• Bladder urothelial carcinoma (BLCA)

• Breast invasive carcinoma (BRCA)

• Cervical squamous cell carcinoma (CESC)

• Cholangiocarcinoma (CHOL)

• Colon adenocarcinoma (COAD)

• Esophageal carcinoma (ESCA)

• Glioblastoma multiforme (GBM)

• Head and neck squamous cell carcinoma (HNSC)

• Kidney renal clear cell carcinoma (KIRC)

• Kidney renal papillary cell carcinoma (KIRP)

• Liver hepatocellular carcinoma (LIHC)

• Lung adenocarcinoma (LUAD)

• Lung squamous cell carcinoma (LUSC)

• Prostate adenocarcinoma (PRAD)

• Rectum adenocarcinoma (READ)

• Stomach adenocarcinoma (STAD)

• Thyroid carcinoma (THCA)

• Uterine corpus endometrial carcinoma (UCEC)

At the protein level, the Clinical Proteomic Tumor Analysis Consortium (CPTAC) database confirms elevated AURKB protein expression in BRCA, HNSC, LIHC, LUAD, PAAD, and UCEC compared to normal tissues .

What is the diagnostic and prognostic value of AURKB in cancer?

AURKB demonstrates exceptional diagnostic value across multiple cancer types:

These findings collectively position AURKB as a potential pan-cancer biomarker with both diagnostic and prognostic utility.

How does AURKB interact with the tumor immune microenvironment?

AURKB demonstrates significant associations with immune components in the tumor microenvironment:

• Immune cell infiltration: Analysis using the TIMER 2.0 database with multiple algorithms (TIMER, EPIC, TIDE) revealed strong correlations between AURKB expression and the infiltration of various immune cell types, including:

  • B cells

  • CD4+ T cells

  • CD8+ T cells

  • Neutrophils

  • Macrophages

  • Dendritic cells

• Immunomodulatory relationships: AURKB expression correlates with numerous immune-related genes involved in:

  • Major histocompatibility complex (MHC) presentation

  • Immune activation pathways

  • Immunosuppressive mechanisms

  • Chemokine signaling networks

• Potential impact on cancer progression: AURKB appears to influence cancer progression through effects on inflammatory and immune-related pathways, suggesting potential implications for immunotherapy response .

What is the relationship between AURKB and genomic instability markers in tumors?

AURKB expression demonstrates significant associations with key genomic instability markers:

• Tumor Mutational Burden (TMB): Spearman correlation analysis reveals significant relationships between AURKB expression and TMB across multiple cancer types .

• Microsatellite Instability (MSI): Similar correlations exist between AURKB expression and MSI status in various tumors .

• Cell cycle regulation: AURKB has a potential impact on cancer progression through its effects on cell cycle regulation, which may contribute to genomic instability through aberrant mitotic processes .

These associations suggest AURKB may either contribute to or be affected by genomic instability in cancer cells, providing important context for interpreting AURKB expression data and its potential mechanistic roles in tumorigenesis.

How do epigenetic modifications influence AURKB expression in cancer?

Epigenetic regulation of AURKB shows tissue-specific patterns that may contribute to its differential expression in cancer:

• Promoter methylation analysis: The UALCAN database demonstrates that AURKB promoter methylation is decreased in multiple cancer types including BLCA, BRCA, HNSC, KIRC, KIRP, READ, LIHC, LUAD, PRAD, THCA, and UCEC compared to normal tissues .

• Tissue-specific patterns: Conversely, PAAD tissues exhibit hypermethylation of the AURKB gene promoter, highlighting the tissue-specific nature of epigenetic regulation .

• Relationship with expression: These differential methylation patterns likely contribute to the varied expression of AURKB observed across cancer types, with hypomethylation generally associated with increased expression.

Understanding these epigenetic mechanisms provides insights into potential regulatory targets and may inform strategies for modulating AURKB expression in research and therapeutic contexts.

What genetic alterations commonly affect the AURKB gene in cancer?

Analysis using the cBioPortal database reveals that AURKB exhibits several types of genetic alterations across cancer types:

• Primary alteration types:

  • Missense mutations

  • Deep deletions

  • Amplifications

• Cancer distribution: AURKB mutations are commonly observed in various cancers including:

  • Diffuse large B-cell lymphoma (DLBC)

  • Sarcoma (SARC)

  • Uterine corpus endometrial carcinoma (UCEC)

  • Liver hepatocellular carcinoma (LIHC)

  • Stomach adenocarcinoma (STAD)

  • And numerous others

• Functional implications: These genetic alterations may affect AURKB function, expression levels, or interactions with other proteins, potentially contributing to its role in cancer development and progression.

Understanding the landscape of genetic alterations affecting AURKB provides critical context for interpreting functional studies and developing therapeutic strategies targeting AURKB or related pathways.

How can researchers resolve contradictory findings regarding AURKB expression or function?

When faced with contradictory findings in AURKB research, consider these methodological approaches:

• Tissue heterogeneity assessment: Implement spatial profiling techniques to evaluate AURKB expression across different regions of tumors, as heterogeneity may explain apparent contradictions.

• Methodological standardization: Carefully document and compare protocols, including:

  • Antibody clones and dilutions

  • Detection methods (chromogenic vs. fluorescent)

  • Quantification approaches (manual vs. digital pathology)

  • Scoring systems and cutoff values

• Patient cohort stratification: Analyze results in the context of:

  • Demographic factors

  • Treatment history

  • Disease stage and grade

  • Molecular subtypes

• Isoform-specific analysis: Ensure primers, probes, and antibodies detect relevant AURKB isoforms, as splice variants may display different expression patterns or functions.

• Post-translational modification consideration: Assess whether phosphorylation or other modifications might affect antibody binding or protein function in different experimental systems.

• Multi-omics integration: Combine protein expression data with transcriptomic, methylation, and mutation analyses to resolve apparent contradictions through a more comprehensive understanding of AURKB regulation.

What emerging research areas are investigating AURKB beyond its classical mitotic functions?

Current frontiers in AURKB research extend beyond its established mitotic roles:

• Non-mitotic functions: Investigating AURKB's potential roles in interphase cells, including possible functions in gene expression regulation and DNA damage response.

• Tumor microenvironment interactions: Exploring how AURKB expression in cancer cells influences surrounding stromal and immune cells, particularly given its associations with immune infiltration .

• Therapeutic resistance mechanisms: Examining whether AURKB contributes to resistance against conventional therapies through promotion of genomic instability or other mechanisms.

• Combination therapy approaches: Investigating whether AURKB inhibition might sensitize cancer cells to other treatments, including immune checkpoint inhibitors, based on its correlations with immune-related pathways .

• Liquid biopsy applications: Evaluating whether AURKB or its downstream targets could serve as circulating biomarkers for cancer detection or monitoring.

How might AURKB analysis be integrated into precision oncology approaches?

AURKB assessment could contribute to precision oncology through multiple avenues:

• Biomarker development: Given its high diagnostic accuracy (AUC > 0.9) in 21 cancer types, AURKB expression could serve as a pan-cancer diagnostic or prognostic biomarker .

• Patient stratification: AURKB expression patterns might identify patient subgroups most likely to benefit from specific therapeutic approaches, including AURKB inhibitors or immunotherapies.

• Therapeutic target validation: The association between AURKB and cancer progression supports its continued investigation as a therapeutic target, particularly in cancers with consistently high expression.

• Companion diagnostics: As AURKB-targeted therapies advance in development, corresponding diagnostic assays could be developed to identify suitable patients.

• Multi-biomarker panels: Integration of AURKB with other molecular markers, including its associations with TMB and MSI, could enhance the precision of cancer classification and treatment selection .