SLC22A11 Antibody

What is SLC22A11 and why is it important in research?

SLC22A11 (OAT4) is involved in sodium-independent transport and excretion of organic anions, many of which are potentially toxic. It is an integral membrane protein primarily expressed in the kidney and placenta, where it plays a protective role by preventing harmful organic anions from reaching the fetus . The protein's role in transporting compounds like estrone sulfate and dehydroepiandrosterone sulfate makes it relevant for research in renal physiology, pharmacology, and toxicology . Its association with gout susceptibility (particularly in different ethnic populations) has also made it an important target for genetic and clinical research .

What types of SLC22A11 antibodies are available for research?

Most commercially available SLC22A11 antibodies are rabbit polyclonal antibodies that target different regions of the protein. These include:

• N-terminal specific antibodies that recognize the amino-terminal region of SLC22A11

• C-terminal specific antibodies that bind to the carboxy-terminal region

• Antibodies recognizing specific amino acid sequences (e.g., AA 40-150)

• Full-length protein antibodies that recognize the entire SLC22A11 protein

The majority are unconjugated primary antibodies suitable for techniques like Western blotting, immunofluorescence, immunohistochemistry, and ELISA, depending on the specific antibody .

Which species reactivity should I consider when selecting an SLC22A11 antibody?

When selecting an SLC22A11 antibody, consider both your experimental system and the antibody's documented reactivity. Common reactivity patterns include:

For cross-species studies, verify sequence homology. For example, some antibodies against human SLC22A11 show limited reactivity with mouse (50%) and rat (52%) orthologs due to sequence differences .

What are the recommended protocols for Western blotting with SLC22A11 antibodies?

For optimal Western blotting with SLC22A11 antibodies:

• Prepare tissue or cell lysates using RIPA buffer with protease inhibitors

• Load 20-30 μg of protein per lane on 10% SDS-PAGE gels

• Transfer to PVDF membrane at 100V for 90 minutes (wet transfer recommended for membrane proteins)

• Block with 5% non-fat milk in TBST for 1 hour at room temperature

• Incubate with primary SLC22A11 antibody at 1:500-1:1000 dilution overnight at 4°C

• Wash 3× with TBST, 10 minutes each

• Incubate with appropriate HRP-conjugated secondary antibody (typically anti-rabbit IgG) at 1:5000 dilution for 1 hour at room temperature

• Wash 3× with TBST, 10 minutes each

• Develop using enhanced chemiluminescence reagents

Expected band size for human SLC22A11 is approximately 62-65 kDa, though this may vary depending on post-translational modifications .

How can I optimize immunofluorescence staining with SLC22A11 antibodies?

For immunofluorescence staining of SLC22A11:

• Fix cells with 4% paraformaldehyde for 15 minutes at room temperature

• Permeabilize with 0.1% Triton X-100 in PBS for 10 minutes

• Block with 1-5% BSA in PBS for 30-60 minutes

• Incubate with SLC22A11 antibody at 1:100-1:200 dilution in blocking buffer overnight at 4°C

• Wash 3× with PBS, 5 minutes each

• Incubate with fluorophore-conjugated secondary antibody at 1:500 dilution for 1 hour at room temperature

• Wash 3× with PBS, 5 minutes each

• Counterstain nucleus with DAPI (1:1000) for 5 minutes

• Mount and visualize

Since SLC22A11 is a membrane protein, you should observe predominantly membrane localization, particularly in polarized epithelial cells. For kidney tissues, SLC22A11 is typically expressed in the apical membrane of proximal tubule cells .

What controls should I include when working with SLC22A11 antibodies?

Include the following controls for rigorous validation:

• Positive control: Kidney or placenta tissue/cell lysates (known to highly express SLC22A11)

• Negative control: Tissues or cells that don't express SLC22A11 (e.g., muscle tissue)

• Technical controls:

  • Primary antibody omission control

  • Isotype control (rabbit IgG at equivalent concentration)

  • Peptide competition/blocking assay (pre-incubate antibody with immunizing peptide)

• Knockdown/knockout validation: Cells with SLC22A11 silenced via siRNA as demonstrated in studies using siSLC22A11 sequences

• Overexpression validation: Cells transfected with SLC22A11 expression plasmid (e.g., PCMV3-SLC22A11)

These controls help confirm antibody specificity and minimize false positive/negative results .

How can I measure SLC22A11 expression at the mRNA level to complement antibody-based protein detection?

For comprehensive analysis of SLC22A11 expression, complement protein detection with mRNA quantification:

• Isolate total RNA using TRIzol reagent following the manufacturer's protocol

• Assess RNA quality via spectrophotometry (A260/A280 ratio ~2.0) and agarose gel electrophoresis (intact 28S and 18S rRNA bands)

• Synthesize cDNA using reverse transcription kit (e.g., Superscript III) with 1 μg of purified total RNA

• Perform quantitative RT-PCR (qRT-PCR) using:

  • Specific primer and probe sets for human SLC22A11

  • Reference gene primers (e.g., GAPDH) for normalization

  • Appropriate thermal cycling conditions (typically 95°C for 10 min followed by 40 cycles of 95°C for 15s and 60°C for 1 min)

• Analyze data using the ΔΔCt method after normalization to endogenous control

This approach allows correlation between protein and mRNA levels, particularly useful when investigating genetic variants or regulatory mechanisms affecting SLC22A11 expression.

What strategies can I use to study genetic variants of SLC22A11 in relation to antibody detection?

When studying SLC22A11 genetic variants:

• Genotyping approaches:

  • Use established SNP panels targeting known SLC22A11 variants (e.g., rs17299124, rs2078267, rs693591)

  • Consider whole gene sequencing to identify novel variants

• Expression analysis:

  • Generate expression constructs containing different SLC22A11 variants

  • Transfect into appropriate cell lines

  • Use antibodies to assess expression levels and subcellular localization

• Functional characterization:

  • Transport assays with fluorescent or radiolabeled substrates

  • Compare transport kinetics between variants

• Antibody considerations:

  • Verify that your antibody's epitope region is not affected by the variants of interest

  • For nonsynonymous variants that alter amino acid sequence, antibody binding efficiency may differ

  • Use multiple antibodies targeting different regions when studying variants

Research has identified multiple nonsynonymous variants in SLC22A11, with several occurring in predicted transmembrane helix regions that may affect protein structure and function .

How can I investigate SLC22A11 in disease contexts using available antibodies?

For disease-oriented SLC22A11 research:

• Tissue expression analysis:

  • Compare SLC22A11 protein levels in normal versus diseased tissues using immunohistochemistry or Western blotting

  • Use tissue microarrays for high-throughput screening across multiple patient samples

• Genetic association studies:

  • Correlate SLC22A11 genetic variants with disease risk (e.g., gout susceptibility)

  • Examine expression differences based on genotype using antibody-based methods

• Functional studies:

  • Investigate how disease conditions alter SLC22A11 localization or function

  • Use cell models mimicking disease states (e.g., inflammation, hyperuricemia)

• Co-localization studies:

  • Perform dual immunofluorescence with SLC22A11 antibodies and markers of disease pathology

  • Analyze trafficking changes in response to disease stimuli

Studies have demonstrated associations between SLC22A11 variants and gout risk in different ethnic populations, with haplotype analysis revealing protective and risk-conferring genetic patterns .

What are common issues with SLC22A11 antibody detection and how can I resolve them?

For membrane proteins like SLC22A11, sample preparation is particularly critical. Consider using mild detergents or specialized membrane protein extraction kits to maintain native conformation .

How do I select the appropriate application method for my SLC22A11 research?

Select the optimal detection method based on your research questions:

For SLC22A11, its membrane localization and domain structure may impact antibody accessibility, particularly in methods where protein conformation is preserved .

How can I validate SLC22A11 antibody specificity in my experimental system?

For comprehensive antibody validation:

• Literature cross-reference:

  • Review published studies using the same antibody

  • Compare detection patterns across different tissues/cells

• Multi-technique confirmation:

  • Verify consistent results across different detection methods

  • Compare results between antibodies targeting different epitopes

• Molecular validation:

  • Perform gene silencing experiments (siRNA/shRNA) and confirm protein reduction

  • Conduct overexpression studies and verify increased signal

• Targeted validation:

  • Use peptide competition assays with the immunizing peptide

  • Verify absence of signal in known negative tissues

• Orthogonal validation:

  • Correlate protein detection with mRNA levels by qRT-PCR

  • Compare results with mass spectrometry data if available

Studies have used approaches like qRT-PCR with specific SLC22A11 primer and probe sets to validate antibody findings, demonstrating correlation between protein and mRNA expression patterns .

How can SLC22A11 antibodies contribute to research on transporter-mediated drug interactions?

SLC22A11 antibodies can facilitate research on drug interactions through:

• Expression monitoring:

  • Quantify SLC22A11 levels in different tissues to predict drug handling

  • Evaluate regulation of expression in response to drug treatments

• Localization studies:

  • Track changes in membrane localization during drug exposure

  • Investigate trafficking mechanisms affecting drug transport

• Structure-function analysis:

  • Identify critical domains for drug binding using epitope-specific antibodies

  • Correlate structural features with transport efficiency

• Clinical translation:

  • Evaluate SLC22A11 expression in patient samples to predict drug response

  • Develop personalized medicine approaches based on transporter profiles

Since SLC22A11 can transport various compounds including estrone sulfate, dehydroepiandrosterone sulfate, and potentially certain medications, antibody-based research can help understand variability in drug disposition and response .

What is the role of SLC22A11 in disease mechanisms based on current antibody-mediated research?

SLC22A11 has been implicated in several pathological conditions:

• Gout and hyperuricemia:

  • Genetic studies have identified associations between SLC22A11 variants and gout risk

  • Different haplotypes show population-specific protective or risk effects

  • Antibody-based studies can clarify how variants affect protein expression and function

• Kidney disorders:

  • As a transporter involved in organic anion handling, SLC22A11 may contribute to kidney injury

  • Expression changes in disease states can be monitored with antibodies

• Pregnancy complications:

  • Placental expression suggests roles in maternal-fetal transport

  • May protect the fetus from potentially harmful compounds

• Cancer research:

  • Some studies have examined SLC family members as potential biomarkers or therapeutic targets in colorectal cancer and other malignancies

  • Expression profiling using antibodies can identify altered transporter patterns

Research has demonstrated ancestral-specific effects of SLC22A11 genetic variants on gout risk, suggesting complex interactions that may influence clinical manifestations .

How do SLC22A11 antibodies compare with antibodies against related transporters?

When designing experiments involving multiple transporters, consider using antibodies raised in different host species to facilitate co-localization studies .

What experimental designs are optimal for investigating SLC22A11 function using antibodies?

For comprehensive functional studies:

• Expression-function correlation:

  • Quantify SLC22A11 expression using antibodies in different cell models

  • Correlate with transport activity using fluorescent or radiolabeled substrates

  • Analyze how expression levels impact functional outcomes

• Regulatory mechanisms:

  • Treat cells with various stimuli (hormones, drugs, cytokines)

  • Monitor changes in SLC22A11 expression and localization using antibodies

  • Determine signaling pathways involved in regulation

• Genetic manipulation approaches:

  • Generate stable cell lines with varied SLC22A11 expression

  • Create cell models expressing different genetic variants

  • Use antibodies to verify expression and localization differences

  • Correlate with functional assays

• In vivo relevance:

  • Compare findings from cell models with tissue samples

  • Analyze SLC22A11 expression in different physiological and pathological states

  • Consider species differences in expression patterns

Studies have employed these approaches to investigate how genetic variants influence SLC22A11 function, particularly in the context of gout susceptibility and urate handling .