AKR4C10 Antibody

What is AKR1B10 and what are its primary biological functions?

AKR1B10 catalyzes the NADPH-dependent reduction of various carbonyl-containing compounds to their corresponding alcohols. It displays strong enzymatic activity toward all-trans-retinal, 9-cis-retinal, and 13-cis-retinal. AKR1B10 plays a critical role in detoxifying dietary and lipid-derived unsaturated carbonyls, such as crotonaldehyde, 4-hydroxynonenal, trans-2-hexenal, trans-2,4-hexadienal and their glutathione-conjugates carbonyls (GS-carbonyls). Importantly, it displays no reductase activity towards glucose, distinguishing it from other AKR family members .

What are the alternative names for AKR1B10 in the scientific literature?

When searching literature or databases, researchers should be aware that AKR1B10 may be referenced under several alternative names including: AKR1B11, Aldo-keto reductase family 1 member B10, ARL-1, Aldose reductase-like, Aldose reductase-related protein, Small intestine reductase, ARP, hARP, and SI reductase . This diversity in nomenclature is important when conducting comprehensive literature reviews.

What types of AKR1B10 antibodies are available for research applications?

Currently, rabbit polyclonal antibodies against AKR1B10 are widely available for research purposes. These antibodies are typically generated against recombinant protein fragments containing sequences within human AKR1B10 amino acids 1-300 or 1-286 . Polyclonal antibodies offer the advantage of recognizing multiple epitopes, potentially increasing detection sensitivity, but may have batch-to-batch variation that should be considered in experimental design.

What techniques are validated for AKR1B10 antibodies in current research?

AKR1B10 antibodies have been validated for several key techniques:

• Western Blot (WB): For quantitative protein expression analysis

• Immunocytochemistry/Immunofluorescence (ICC/IF): For cellular localization studies

• Immunohistochemistry (IHC): For tissue expression pattern analysis

The choice of technique should be guided by experimental questions and appropriate validation controls for each application .

What species reactivity can be expected with commercial AKR1B10 antibodies?

Commercial antibodies have demonstrated reactivity with human samples across multiple applications. Some antibodies also react with mouse and rat samples, though cross-reactivity should be verified experimentally for each specific antibody . When studying AKR1B10 in animal models, it's critical to confirm antibody specificity due to potential sequence differences across species.

What are the optimal conditions for Western blot detection of AKR1B10?

For optimal Western blot results with AKR1B10 antibodies:

• Sample preparation: Use 30 μg of whole cell lysate

• Separation: Run samples on 10% SDS-PAGE

• Primary antibody: Dilute at 1:1000 concentration

• Detection: Use appropriate secondary antibodies (anti-rabbit IgG)

Validation data shows successful detection in multiple cell lines including MOLT4 and Raji cells . Researchers should include positive controls such as liver or small intestine tissue extracts where AKR1B10 is naturally expressed.

What considerations are important for immunohistochemical detection of AKR1B10?

For immunohistochemistry applications:

• Sample preparation: Use paraffin-embedded tissue sections

• Antigen retrieval: Heat-induced epitope retrieval is typically necessary

• Antibody dilution: Start with 1:500 dilution and optimize as needed

• Controls: Include positive controls (such as OVCAR3 xenograft tissue) and negative controls (primary antibody omission)

• Counterstaining: Use hematoxylin for nuclear visualization

Quantification should follow standardized scoring systems that account for both staining intensity and percentage of positive cells.

How should researchers approach knockdown experiments to study AKR1B10 function?

When performing AKR1B10 knockdown experiments:

• Vector selection: Use lentiviral shRNA systems for stable knockdown

• Transduction protocol: Apply lentivirus at approximately 100 μl/ml (1×10^9 TU/ml) when cell confluency reaches 60-70%

• Validation: Confirm knockdown efficiency at both mRNA and protein levels

• Controls: Include scrambled shRNA controls

• Phenotypic assays: Assess effects on proliferation (CCK-8, colony formation), migration (Transwell, wound healing), and invasion as appropriate for your research question

Multiple shRNA constructs targeting different regions of AKR1B10 are recommended to rule out off-target effects.

How does AKR1B10 expression vary across different cancer types?

AKR1B10 expression shows significant variation across cancer types. Analysis of GDC-TCGA and ICGC databases reveals that AKR1B10 is significantly upregulated in several cancers, with particularly notable expression in liver cancer . The Human Protein Atlas database provides immunohistochemical images that demonstrate this differential expression pattern across tumor samples . When analyzing novel cancer samples, researchers should normalize expression data appropriately, typically using log2(TPM+1) transformation for RNA-seq data.

What signaling pathways interact with AKR1B10 in cancer progression?

AKR1B10 interacts with several key oncogenic pathways:

These interactions suggest that AKR1B10 functions as a hub protein affecting multiple cancer hallmarks simultaneously.

How does AKR1B10 influence cancer cell proliferation and metastasis?

Research demonstrates that AKR1B10 significantly affects cancer cell behavior:

• Proliferation: AKR1B10 knockdown in Huh7 liver cancer cells reduces proliferation capacity

• Migration and Invasion: Diminished AKR1B10 expression decreases both migration and invasion potential

• EMT: AKR1B10 regulates epithelial-mesenchymal transition by modulating key markers including E-cadherin, N-cadherin, vimentin, and Twist1

DepMap analysis confirms that AKR1B10 activity influences HCC cell proliferation, migration, and invasion capabilities . These findings suggest AKR1B10 as a potential therapeutic target.

How can AKR1B10 antibodies be used in combination with other detection methods?

For comprehensive analysis, researchers should consider combining antibody-based methods with other detection approaches:

• Transcriptomic validation: Correlate protein detection with RNA-seq or qPCR data

• Mass spectrometry: Confirm antibody specificity and identify post-translational modifications

• Activity assays: Combine expression analysis with enzymatic activity measurements to correlate abundance with function

• Single-cell analysis: Evaluate heterogeneity of AKR1B10 expression within tumor populations

This multi-modal approach provides more robust evidence than relying solely on antibody-based detection.

What are effective strategies for studying AKR1B10's role in therapy resistance?

To investigate AKR1B10's potential role in therapy resistance:

• Compare expression in paired pre- and post-treatment samples

• Generate resistant cell lines and analyze AKR1B10 expression changes

• Perform knockdown/overexpression in resistant cells to assess functional impact

• Combine AKR1B10 inhibition with standard therapies to test for synergistic effects

• Analyze patient cohorts stratified by AKR1B10 expression levels for treatment response differences

Recent developments in antibody-based therapies for resistant cancers, such as AHA-1031 , provide conceptual frameworks for developing AKR1B10-targeted approaches.

How can researchers investigate AKR1B10's interactions with immune cells in the tumor microenvironment?

This emerging research area requires sophisticated experimental approaches:

• Multiplex immunofluorescence: Co-stain for AKR1B10 and immune cell markers

• Spatial transcriptomics: Analyze co-expression patterns of AKR1B10 and immune genes

• Co-culture experiments: Assess how AKR1B10-expressing cancer cells affect immune cell function

• In vivo models: Evaluate tumor growth and immune infiltration in AKR1B10-manipulated xenografts

• Correlation with immunotherapy response: Analyze whether AKR1B10 expression predicts response to immune checkpoint inhibitors

Novel antibody-based immunotherapies like those targeting NK cells might provide insights into potential therapeutic strategies involving AKR1B10.

What statistical approaches are recommended for analyzing AKR1B10 expression in cancer datasets?

For robust statistical analysis:

How should conflicting results about AKR1B10's role in different cancer contexts be interpreted?

When facing contradictory findings:

• Context specificity: Consider that AKR1B10 may have opposite roles in different cancer types

• Methodological differences: Evaluate antibody specificity, detection methods, and scoring systems

• Experimental models: Compare in vitro vs. in vivo vs. clinical sample results

• Cancer stage: Analyze whether AKR1B10's role changes during cancer progression

• Genetic background: Consider how the underlying mutational landscape affects AKR1B10 function

What future directions should researchers consider for AKR1B10 antibody applications?

Emerging research directions include:

• Development of more specific monoclonal antibodies targeting diverse epitopes

• Therapeutic antibody conjugates targeting AKR1B10-expressing tumors

• Liquid biopsy applications to detect circulating AKR1B10 protein

• Single-cell analysis of AKR1B10 heterogeneity in tumors

• Combination biomarker approaches incorporating AKR1B10 with other markers

The development of novel antibody-based technologies like those seen in immunotherapy-resistant cancer research might eventually be applied to AKR1B10-targeted therapeutics.