ALDH1B1 Antibody

What tissue types express ALDH1B1 and can be detected using ALDH1B1 antibodies?

ALDH1B1 expression has been detected in multiple tissues through antibody-based approaches. Based on immunohistochemistry data, ALDH1B1 is prominently expressed in:

• Pancreatic buds during early development (e9.5 stage) with expression patterns similar to PDX1

• Colon tissue, particularly in the stem cell compartment

• Human adenocarcinomas of colon, lung, breast, and ovary

• Testicular tissue

• Liver tissue

• Brain tissue (mouse)

For optimal detection in these tissues, immunohistochemistry protocols typically recommend antigen retrieval with TE buffer at pH 9.0, although citrate buffer at pH 6.0 may also be used as an alternative . When studying developmental expression patterns, combining in situ hybridization with immunofluorescence provides valuable confirmation of antibody specificity and expression localization .

What are the recommended applications and dilutions for ALDH1B1 antibodies?

Based on validated research protocols, ALDH1B1 antibodies can be used in multiple applications with specific dilution ranges:

The optimal dilution should be determined experimentally for each specific application and sample type . For example, when performing IHC on paraffin-embedded human breast cancer tissue, a 1:100 dilution has been successfully employed .

How can researchers verify the specificity of ALDH1B1 antibodies?

Antibody specificity verification is crucial for reliable research. For ALDH1B1, multiple approaches have been documented:

• Western immunoblotting against recombinant proteins: Testing reactivity against multiple ALDH family members. For example, some anti-ALDH1B1 antibodies show strong immunoreactivity toward ALDH1B1 but may exhibit cross-reactivity with ALDH1A1 protein .

• Double in situ/immunofluorescence validation: Combined ALDH1B1 in situ hybridization with ALDH1B1 immunofluorescence can confirm that all ALDH1B1+ cells (by antibody detection) also express ALDH1B1 mRNA .

• Knockout validation: Using tissues from ALDH1B1 knockout mice provides the gold standard for antibody validation. Studies with ALDH1B1 knockout mice have confirmed no compensatory changes in the expression of related ALDH2 or ALDH1A1 genes, making these models valuable for antibody validation .

• Immunogen specificity: Examining the antibody's immunogen sequence for potential cross-reactivity with other ALDH family members. Some antibodies are raised against specific peptide fragments (e.g., amino acids 353-411 of human ALDH1B1) and purified through antigen-bound affinity columns .

What are the optimal tissue preparation methods for ALDH1B1 immunohistochemistry?

For reliable ALDH1B1 detection in tissues, the following protocol has been validated:

• Fixation and sectioning: Paraffin-embedded tissue sections (5-μm) should be used.

• Deparaffinization and rehydration: Followed by heat-induced epitope retrieval.

• Antigen retrieval: Use Retrieval Solution (e.g., TE buffer pH 9.0) at 90°C for 10 minutes. Alternatively, citrate buffer (pH 6.0) can be used, though potentially with different results .

• Protein blocking: Apply Protein Blocker before antibody incubation.

• Primary antibody incubation: Apply anti-ALDH1B1 antibody at appropriate dilution (e.g., 1:750 for polyclonal anti-human ALDH1B1) in Protein Blocker for 60 minutes at room temperature.

• Secondary antibody: Apply HRP-conjugated secondary antibodies (1:500) for 10-20 minutes at room temperature.

• Visualization: Incubate with DAB for 10 minutes, followed by counterstaining with Hematoxylin .

For developing embryonic tissues, whole mount in situ hybridization combined with tyramide signal amplification has been successfully employed to detect ALDH1B1 expression in early developmental stages .

How can ALDH1B1 antibodies be used to investigate cancer stem cells?

ALDH1B1 has emerged as a potential biomarker for cancer stem cells, particularly in colorectal cancer. Research methodologies include:

• Comparative expression analysis: Studies have found 5.6-fold higher expression scores for ALDH1B1 in cancerous tissues compared to ALDH1A1, with 39 out of 40 colonic cancer specimens showing positive staining for ALDH1B1 with a mean intensity of 2.8 ± 0.5 .

• Prognostic correlation studies: High ALDH1B1 expression has been correlated with poor prognosis in colorectal cancer patients. Immunohistochemistry scoring systems that assess both intensity (1-3 scale) and extensiveness (% of total cancer cells) can be employed to quantify expression levels .

• Immune cell infiltration analysis: ALDH1B1 expression has been correlated with various immune cell markers, particularly in head and neck squamous cell carcinoma (HNSC) and stomach adenocarcinoma (STAD). The relationship between ALDH1B1 expression and six types of immune cell infiltration (B cells, CD8+ T cells, CD4+ T cells, macrophages, DCs, and neutrophils) provides insights into the tumor microenvironment .

The following correlation data illustrates ALDH1B1's relationship with immune markers in HNSC and STAD:

*** indicates p < 0.0001

What is the role of ALDH1B1 in cancer detection and how can antibodies facilitate biomarker development?

ALDH1B1 shows promise as a biomarker for early cancer detection, particularly for colorectal cancer and advanced adenoma:

• Autoantibody detection: ALDH1B1 autoantibodies have demonstrated detection value for colorectal cancer (CRC) and advanced adenoma with area under the curve (AUC) values of 0.70 and 0.74, respectively .

• ELISA development: For detecting ALDH1B1 autoantibodies in serum, ELISA protocols have been developed that show sensitivity of 75.68% and specificity of 63.06% for CRC, and sensitivity of 62.31% and specificity of 73.87% for advanced adenoma .

• Biomarker panels: Combining ALDH1B1 autoantibodies with other markers (UQCRC1, CTAG1, and CENPF) improves performance with an AUC of 0.79 for detecting both CRC and advanced adenomas .

• Correlation with tumor stage: Research protocols can examine ALDH1B1 expression across different cancer stages using tissue microarrays and standardized scoring systems that account for both staining intensity and extensiveness .

How should researchers address cross-reactivity concerns with ALDH1B1 antibodies?

Cross-reactivity with other ALDH family members is a significant concern due to sequence homology. Strategies to address this include:

• Pre-validation of antibody specificity: Test antibody reactivity against multiple recombinant ALDH proteins. For example, some anti-ALDH1B1 antibodies show strong reactivity to ALDH1B1 but some cross-reactivity to ALDH1A1 .

• Use of appropriate controls: Include ALDH1B1 knockout samples when available. Global knockout mouse strains for ALDH1B1 have been generated and show no compensatory changes in related ALDH genes .

• Comparative analysis: When using polyclonal antibodies, compare results with monoclonal antibodies targeting different epitopes to confirm specificity.

• Immunogen verification: Select antibodies raised against unique regions of ALDH1B1 with minimal homology to other ALDH family members .

• Multiple detection methods: Confirm findings using both protein detection (antibody-based) and mRNA detection methods (e.g., in situ hybridization) to validate expression patterns .

How can researchers optimize ALDH1B1 antibody-based detection in challenging tissue samples?

For difficult-to-analyze samples, optimize protocols through:

• Refined antigen retrieval: For tissues with high fixative cross-linking, extend heat-induced epitope retrieval time or test alternative buffers. Compare TE buffer pH 9.0 with citrate buffer pH 6.0 to determine optimal conditions .

• Signal amplification: For low expression levels, implement tyramide signal amplification systems, which have been successfully used with ALDH1B1 in situ hybridization in embryonic tissues .

• Blocking optimization: For tissues with high background, increase blocking duration or concentration, or try alternative blocking reagents beyond standard protein blockers.

• Antibody titration: Perform detailed dilution series experiments to identify the optimal antibody concentration that maximizes specific signal while minimizing background for each application and tissue type .

• Combined approaches: For ambiguous results, integrate multiple detection methods. For example, RNA-scope in situ hybridization can be used alongside immunohistochemistry to validate expression patterns in challenging samples.

How can ALDH1B1 antibodies contribute to understanding the link between alcohol metabolism and cancer?

ALDH1B1 has dual relevance in alcohol metabolism and cancer development. Research approaches include:

• Functional analysis in knockout models: ALDH1B1 knockout mice exhibit approximately 40% greater accumulation of blood acetaldehyde following acute ethanol exposure, confirming ALDH1B1's role in acetaldehyde clearance in vivo .

• Polymorphism studies: Several functional polymorphisms of the ALDH1B1 gene have been identified in human populations. Alcohol hypersensitivity has been observed in carriers of the ALDH1B1*2 variant, which metabolizes acetaldehyde at a slower rate than wild-type ALDH1B1 .

• Cancer risk correlation: Investigation of ALDH1B1 polymorphisms in relation to cancer susceptibility, particularly in populations with varying alcohol consumption patterns, can be facilitated by specific antibodies that recognize variant forms of the protein.

• Mechanistic pathway studies: Using ALDH1B1 antibodies to track protein expression and localization in response to alcohol exposure provides insights into the mechanistic link between alcohol metabolism and carcinogenesis.

What is the potential of ALDH1B1 as a therapeutic target, and how can antibodies facilitate drug development research?

ALDH1B1's association with cancer stem cells and poor prognosis suggests therapeutic potential:

What novel ALDH1B1 antibody applications are emerging in single-cell analysis of tumor heterogeneity?

With advances in single-cell technologies, ALDH1B1 antibodies offer potential for deeper understanding of tumor heterogeneity:

• Single-cell sorting and analysis: ALDH1B1 antibodies can be used in flow cytometry and cell sorting to isolate rare subpopulations of ALDH1B1-expressing cells within tumors for subsequent molecular characterization.

• Spatial transcriptomics integration: Combining ALDH1B1 immunofluorescence with spatial transcriptomics can reveal spatial relationships between ALDH1B1-expressing cells and other cell types within the tumor microenvironment.

• Lineage tracing studies: In developmental biology and cancer research, ALDH1B1 antibodies can help track cell lineages, particularly in the context of stem/progenitor cells in pancreatic development where ALDH1B1 shows specific expression in pancreatic buds .

• Multi-parameter imaging: Integration of ALDH1B1 antibodies into multiplexed immunofluorescence panels alongside immune cell markers can provide comprehensive visualization of tumor-immune interactions, particularly important given the correlations observed between ALDH1B1 expression and immune cell infiltration markers .

How can researchers integrate ALDH1B1 antibody data with other omics approaches for comprehensive cancer characterization?

Multi-omics integration strategies include:

• Proteogenomic correlation: Correlating ALDH1B1 protein expression (detected by antibodies) with genetic alterations and transcriptomic profiles can reveal regulatory mechanisms and functional consequences of ALDH1B1 dysregulation.

• Epigenetic modification analysis: Investigating how epigenetic modifications affect ALDH1B1 expression by combining chromatin immunoprecipitation studies with antibody-based protein detection provides insights into regulatory mechanisms.

• Metabolomic integration: Given ALDH1B1's enzymatic function in aldehyde metabolism, correlating its expression with metabolomic profiles can elucidate functional consequences of its dysregulation in cancer metabolism.

• Systems biology approaches: Integrating ALDH1B1 antibody data into pathway and network analyses can position ALDH1B1 within broader cellular signaling networks and identify potential therapeutic intervention points.

• Clinical outcome correlation: Combining ALDH1B1 expression data from antibody-based studies with clinical outcome data enables identification of prognostic signatures and potential therapeutic targets, particularly important given ALDH1B1's association with poor prognosis in several cancer types .