SULT1A2 Antibody

What is SULT1A2 and why is it significant in research?

SULT1A2 belongs to the cytosolic sulfotransferase family of phase II detoxification enzymes. This protein catalyzes the sulfate conjugation of various compounds including hormones, neurotransmitters, drugs, and xenobiotics . Its significance in research stems from its potential role in chemical carcinogenesis, as studies with recombinant enzymes have demonstrated that SULT1A2 can catalyze the bioactivation of several procarcinogens . Recent research has also revealed its potential role as a biomarker in bladder cancer, where its expression correlates with tumor staging and patient prognosis .

How can researchers detect SULT1A2 expression in tissue samples?

Detection of SULT1A2 expression can be accomplished through multiple complementary techniques:

• Immunohistochemistry (IHC): Using specific antibodies like HPA051051, researchers can detect SULT1A2 protein in formalin-fixed, paraffin-embedded (FFPE) tissues. In bladder cancer research, IHC has successfully demonstrated cytoplasmic localization of SULT1A2 .

• Western blot analysis: This technique allows for semi-quantitative assessment of SULT1A2 protein expression in tissue or cell lysates, providing information about protein size and relative abundance .

• Reverse transcription-quantitative PCR (RT-qPCR): For examining SULT1A2 at the mRNA expression level, RT-qPCR provides a reliable method to quantify transcript levels .

• Database analysis: Utilizing public databases such as The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) can provide complementary expression data across various tissue types and disease states .

What validation steps are essential when using SULT1A2 antibodies?

When using SULT1A2 antibodies, particularly for studies where expression levels may be controversial, rigorous validation is critical:

• Antibody specificity testing: The antibody should be tested against recombinant SULT1A2 and other SULT family members to confirm specificity. For example, researchers have developed antibodies directed against regions of human SULT1A2 that differ from other known sulfotransferase isoforms .

• Positive and negative controls: Include tissues or cell lines with known SULT1A2 expression levels. Research has shown that certain bladder cancer cell lines exhibit SULT1A2 expression and can serve as positive controls .

• Multiple detection methods: Corroborate antibody-based detection with orthogonal methods such as RT-qPCR for mRNA expression .

• Blocking peptide controls: Consider using specific blocking peptides to confirm binding specificity in applications like Western blotting and immunohistochemistry.

How should researchers address the controversy regarding SULT1A2 protein expression?

To address this controversy, researchers should:

• Employ multiple detection methods: Combine protein detection (IHC, Western blot) with mRNA analysis (RT-qPCR) to provide comprehensive expression data .

• Investigate tissue-specific expression: Focus on tissues where expression has been reported, such as bladder cancer tissues, rather than assuming consistent expression across all tissue types .

• Consider pathological states: SULT1A2 expression may be induced under certain physiological or pathological conditions. Research has shown significantly higher expression in bladder cancer compared to normal bladder tissues .

• Examine splice variants: Investigate potential splice variants that may affect protein expression and function, particularly as previous studies have suggested splicing defects .

What methodological approaches best characterize SULT1A2's role in carcinogenesis?

To effectively characterize SULT1A2's role in carcinogenesis, researchers should implement:

• Functional studies: Overexpression and knockdown experiments in relevant cell lines to assess effects on proliferation, invasion, and other cancer-related phenotypes.

• Pathway analysis: Gene set enrichment analysis (GSEA) has revealed that SULT1A2 is associated with cancer-related pathways including PI3K-Akt signaling, MAPK signaling, and pathways in cancer .

• Disease ontology analysis: This approach has demonstrated that bladder cancer is among the diseases most closely associated with SULT1A2 .

• Gene Ontology (GO) analysis: Research has shown SULT1A2 variation in bladder cancer results in changes to biosynthetic processes, regulation of metabolic processes, nuclear functions, and DNA binding .

How can SULT1A2 expression patterns be correlated with clinical outcomes?

To establish clinically relevant correlations with SULT1A2 expression:

What are the key challenges in SULT1A2 detection and quantification?

Researchers face several challenges when detecting and quantifying SULT1A2:

• Low expression levels: If SULT1A2 is expressed as protein, levels may be very low in certain tissues, requiring sensitive detection methods .

• Family member similarity: High sequence homology among SULT family members necessitates highly specific antibodies to avoid cross-reactivity .

• Tissue-specific expression: Expression patterns may vary significantly between tissues and disease states, requiring careful selection of positive controls .

• Potential splice variants: Alternative splicing may affect detection depending on the antibody epitope targeted .

What optimization strategies improve SULT1A2 immunohistochemistry results?

To optimize SULT1A2 immunohistochemistry:

• Antigen retrieval optimization: Test different antigen retrieval methods (heat-induced vs. enzymatic) and buffer compositions to maximize epitope exposure.

• Antibody titration: Determine optimal antibody concentration through serial dilutions to achieve specific staining with minimal background.

• Incubation conditions: Optimize temperature and duration for both primary and secondary antibody incubations.

• Signal amplification: Consider using signal amplification systems for low-abundance detection, particularly in tissues where expression may be limited.

• Counterstaining optimization: Select appropriate counterstains to facilitate identification of subcellular localization, as SULT1A2 has been found prominently in the cytoplasm .

How should researchers interpret contradictory findings about SULT1A2 expression?

When encountering contradictory literature regarding SULT1A2 expression:

• Consider methodological differences: Earlier studies using certain antibodies may have had limited sensitivity compared to newer detection methods .

• Evaluate tissue specificity: Expression may be restricted to certain tissues or induced under specific conditions. Recent research found high expression in bladder cancer but not necessarily in all tissue types .

• Assess pathological context: Expression patterns differ between normal and cancerous tissues and between cancer stages, with higher expression in non-muscle invasive compared to muscle invasive bladder cancer .

• Examine threshold-dependent effects: Research suggests SULT1A2 may have different roles at different expression thresholds, potentially explaining contradictory findings .

What pathways and biological processes should be analyzed in SULT1A2 functional studies?

Based on research findings, several key pathways and processes warrant investigation:

• Cancer-related signaling pathways: The PI3K-Akt signaling pathway, MAPK signaling pathway, and general pathways in cancer have been negatively associated with SULT1A2 in bladder cancer .

• Biosynthetic and metabolic processes: SULT1A2 variation in bladder cancer affects biosynthetic and metabolic processes .

• Nuclear functions and DNA binding: GO analysis has demonstrated SULT1A2's association with nuclear functions and DNA binding .

• Human papillomavirus (HPV) infection pathways: Research has shown association between SULT1A2 and HPV infection pathways .

What experimental design considerations are important for SULT1A2 functional studies?

When designing experiments to investigate SULT1A2 function:

• Multiple cell line models: Use diverse cell lines to account for heterogeneity in SULT1A2 expression and function.

• Complementary approaches: Combine overexpression and knockdown/knockout experiments to provide comprehensive functional insights.

• Physiologically relevant conditions: Consider inducible expression systems to mimic the potential conditional expression of SULT1A2 in vivo.

• In vivo validation: After in vitro studies, validate findings in appropriate animal models when possible.

• Comprehensive endpoint analysis: Measure multiple cellular phenotypes (proliferation, migration, invasion, apoptosis) to fully characterize SULT1A2's functional impact .