SLC22A18AS Antibody

What is SLC22A18AS and why is it significant for research?

SLC22A18AS (solute carrier family 22 member 18 antisense) is part of a sense-antisense gene pair located at human chromosome segment 11p15.5. This gene is paternally imprinted, meaning paternal alleles are silenced while maternal alleles are expressed. The significance of SLC22A18AS lies in its genomic relationship with SLC22A18, where they share 31 bp in their 5′ regions in divergent orientations (between the first exon of SLC22A18AS and the second exon of SLC22A18) . Recent research has highlighted the role of SLC22A18AS in various cancers, particularly non-small cell lung cancer (NSCLC), where its expression is associated with disease progression and patient survival .

What are the optimal dilution ratios for different SLC22A18AS antibody applications?

The optimal dilution varies by application and specific antibody formulation:

These recommendations serve as starting points; researchers should perform optimization experiments for their specific samples and conditions. Antibody performance is sample-dependent, and titration is recommended in each testing system to obtain optimal results .

What is the recommended protocol for immunoprecipitation using SLC22A18AS antibodies?

For immunoprecipitation of SLC22A18AS:

• Prepare protein lysates from your cells of interest (e.g., HepG2 cells have been validated)

• Use 0.5-4.0 μg of antibody per 1.0-3.0 mg of total protein lysate

• Follow standard IP protocols with appropriate washing and elution steps

• Perform western blot analysis to confirm successful immunoprecipitation

Note that detailed IP protocols specific to SLC22A18AS antibody 13475-1-AP are available from manufacturers and should be consulted for optimal results .

How can researchers address non-specific binding issues with SLC22A18AS antibodies?

Non-specific binding can be minimized through several approaches:

• Optimization of blocking conditions: Use 3-5% BSA or 5% non-fat milk in TBS-T for western blots

• Titration of antibody concentration: Begin with manufacturer recommendations and adjust as needed

• Inclusion of appropriate controls: Always include negative controls (samples without target protein) and positive controls (validated samples with known SLC22A18AS expression)

• Cross-reactivity assessment: The antibody is designed to recognize SLC22A18AS only, not SLC22A18, despite their genomic relationship

For immunohistochemistry applications, antigen retrieval methods should be optimized, as SLC22A18AS detection has been validated in human colon carcinoma tissues .

What storage conditions are critical for maintaining SLC22A18AS antibody functionality?

Proper storage is crucial for antibody stability and performance:

Aliquoting is unnecessary for -20°C storage of glycerol-containing formulations. For working dilutions, store at 2-8°C for up to one month. Centrifuge briefly prior to opening the vial to collect solution at the bottom .

How does DNA methylation affect SLC22A18AS expression, and what implications does this have for experimental design?

Research has demonstrated that SLC22A18AS expression is regulated by promoter methylation, particularly in cancer contexts. In NSCLC, SLC22A18AS is hypomethylated, leading to overexpression of the gene . This has significant implications for experimental design:

• Methylation analysis: Consider including methylation status assessment when studying SLC22A18AS expression

• Demethylating agents: Treatment with ademetionine has been shown to reverse gene expression profiles in vitro

• Interpretation complexity: Expression patterns may reflect both genetic and epigenetic regulation

When designing experiments involving SLC22A18AS, researchers should consider the methylation status of their experimental models to properly interpret results, especially in cancer-related studies .

What is the relationship between SLC22A18AS and its sense partner SLC22A18, and how might this impact antibody selection?

The SLC22A18/SLC22A18AS sense-antisense pair exhibits a complex relationship with important experimental considerations:

• Genomic overlap: These genes share 31 bp in their 5′ regions between the first exon of SLC22A18AS and the second exon of SLC22A18

• Co-expression patterns: Studies have shown a significant positive correlation between SLC22A18 and SLC22A18AS expression (r = 0.641; p < 0.001) in NSCLC samples

• Functional relationship: Both genes influence similar cancer phenotypes, suggesting potential regulatory interactions

When selecting antibodies, ensure they specifically recognize SLC22A18AS and not SLC22A18. Most commercial antibodies (e.g., 13475-1-AP) are designed to recognize SLC22A18AS only . Cross-reactivity testing is advisable, particularly in experimental contexts where both proteins may be present.

How does SLC22A18AS expression correlate with cancer progression and prognosis?

Multiple studies have demonstrated significant associations between SLC22A18AS expression and cancer outcomes:

These findings suggest SLC22A18AS antibodies may have value in prognostic assessments and translational research aimed at stratifying patients for treatment approaches .

What functional evidence exists for SLC22A18AS's role in tumorigenesis, and how can antibodies help elucidate these mechanisms?

Functional studies using gene knockdown approaches have provided evidence for SLC22A18AS's role in cancer:

• Cell proliferation: SiRNA-mediated silencing of SLC22A18AS significantly impaired cell proliferation in both adenocarcinoma and squamous cell carcinoma cell lines

• Cancer phenotypes: SLC22A18AS overexpression appears to promote tumor cell proliferation, suggesting an oncogenic role

• Molecular mechanisms: SLC22A18AS's function may be linked to membrane transport processes, as it is part of the solute carrier family

SLC22A18AS antibodies can help elucidate these mechanisms through:

• Immunoprecipitation to identify protein interaction partners

• Immunofluorescence to determine subcellular localization

• ChIP assays to investigate chromatin interactions

• Immunohistochemistry to assess expression patterns in different tissue contexts

These approaches can help map the molecular networks through which SLC22A18AS influences cancer progression .

How might SLC22A18AS variants impact antibody binding and experimental outcomes?

Current research has identified SLC22A18 variants that affect protein expression and function, raising questions about potential SLC22A18AS variants. While specific SLC22A18AS variants are less characterized than those of SLC22A18, researchers should consider:

• Epitope location: Confirm whether your antibody's epitope falls within regions of potential variation

• Western blot validation: Observed molecular weights of 30 kDa and 65 kDa have been reported for SLC22A18AS , and variation from these may indicate variant forms

• Cross-population considerations: If working with samples from diverse populations, consider potential genetic variation that might affect antibody binding

For SLC22A18, missense variants (p.Ala6Thr, p.Arg12Gln, and p.Arg86His) have been shown to have significantly lower cell expression than wild type . Similar phenomena might occur with SLC22A18AS variants, potentially affecting antibody detection sensitivity.

What are the implications of using SLC22A18AS antibodies in combination with other biomarkers for cancer research?

Multimarker approaches incorporating SLC22A18AS may enhance diagnostic and prognostic capabilities:

• Complementary biomarkers: Consider pairing SLC22A18AS with other cancer biomarkers for enhanced specificity and sensitivity

• Pathway analysis: SLC22A18AS functions within broader networks of transporters and regulatory proteins; combine with markers of related pathways

• Multiparametric imaging: In immunohistochemistry applications, co-staining with other markers can provide spatial context for SLC22A18AS expression

Researchers developing such approaches should carefully validate antibody compatibility for multiplex applications, as buffer conditions and detection methods may require optimization when working with multiple antibodies simultaneously .

What controls should be implemented when using SLC22A18AS antibodies for studying imprinted gene regulation?

When investigating imprinted gene regulation with SLC22A18AS antibodies, several controls are essential:

• Allele-specific expression analysis: Given SLC22A18AS's imprinted status, controls that distinguish maternal and paternal allele expression are critical

• Methylation controls: Include methylated and demethylated DNA controls when correlating methylation with antibody-detected protein levels

• Tissue-specific reference samples: Expression patterns vary by tissue type, so appropriate reference tissues should be included

• Knockdown validation: siRNA or shRNA against SLC22A18AS can verify antibody specificity

Additionally, when studying the relationship between SLC22A18AS and SLC22A18, separate antibody validation for each target is necessary to ensure specific detection of the intended protein .

What are the optimal experimental conditions for studying SLC22A18AS in different subcellular compartments?

SLC22A18AS has been detected in different subcellular locations, requiring specific experimental approaches:

• Membrane localization: For plasma membrane studies, surface biotinylation assays combined with SLC22A18AS antibodies have been effective

• Cytoplasmic detection: Immunohistochemical analysis has detected SLC22A18AS protein in the cytoplasm of human colon carcinoma cells

• Compartment fractionation: When performing subcellular fractionation, include markers for each compartment (e.g., Na⁺/K⁺ ATPase for plasma membrane, calnexin for ER)

For immunofluorescence applications, appropriate fixation methods are critical. Paraformaldehyde fixation (4%, 15 minutes) followed by permeabilization with 0.1% Triton X-100 has been successfully used for SLC22A18-related studies and may be applicable to SLC22A18AS detection as well .

How should researchers interpret discrepancies between SLC22A18AS antibody signals and mRNA expression data?

When faced with discrepancies between protein detection via antibodies and mRNA expression data for SLC22A18AS, consider several explanations:

• Post-transcriptional regulation: Studies with SLC22A18 variants revealed comparable mRNA levels despite significant protein expression differences, suggesting post-transcriptional mechanisms

• Protein degradation pathways: Treatment with MG132 (proteasomal proteolysis inhibitor) and bafilomycin A₁ (lysosomal degradation inhibitor) has restored expression levels of SLC22A18 variants , suggesting similar mechanisms might affect SLC22A18AS

• Antibody sensitivity thresholds: Protein levels below antibody detection limits may still show mRNA expression

• Technical factors: Sample preparation differences between protein and RNA analyses can contribute to apparent discrepancies

Methodologically, researchers should consider:

• Parallel qPCR and western blot analyses from the same samples

• Proteasome and lysosome inhibitor treatments to assess protein stability

• Polysome profiling to assess translational efficiency

These approaches can help determine whether discrepancies reflect biological regulation or technical limitations .

What bioinformatic resources can complement SLC22A18AS antibody studies?

Several bioinformatic resources can enhance interpretation of SLC22A18AS antibody-based research:

Integration of antibody-based experimental data with these resources can provide a more comprehensive understanding of SLC22A18AS biology and improve interpretation of experimental results in broader biological contexts .