SLCO1C1 Antibody

What is SLCO1C1 and why is it significant in neurological research?

SLCO1C1 (solute carrier organic anion transporter family member 1C1) is a critical protein involved in thyroxine (T4) transport in the brain. This transporter facilitates the entry of T4 into the adult brain, where it is subsequently converted to 3,5,3'-triiodothyronine (T3) . In adults, SLCO1C1 expression is primarily localized to two brain barrier structures: the blood-brain barrier (BBB) and choroid plexus . The protein has a molecular mass of approximately 78.7 kilodaltons . The significance of SLCO1C1 in neurological research stems from its essential role in thyroid hormone metabolism, which is crucial for proper brain development and function. Disruptions in this transport system may contribute to various neurological disorders related to thyroid hormone dysregulation.

What alternative nomenclature exists for SLCO1C1 in scientific literature?

When conducting literature searches related to SLCO1C1, researchers should be aware of several alternative names for this protein:

• OATP1C1

• OATP-F

• OATP14

• OAT-RP-5

• OATP-14

This diverse nomenclature reflects the evolution of our understanding of this transporter family. Using multiple search terms encompassing these alternative names is essential for comprehensive literature reviews in this field.

How does SLCO1C1 expression differ between developmental stages and adult brain?

SLCO1C1 exhibits dynamic expression patterns that change significantly during development. While adult expression is primarily restricted to the BBB and choroid plexus, developmental expression is more widespread. Studies using Slco1c1-Cre transgenic mice crossed with Rosa26 reporter mice revealed transient Slco1c1 expression during brain development in neurons of various brain structures, including cortical layer 2/3 and the hippocampus . This suggests that SLCO1C1 may play additional roles during brain development beyond its established function in the adult brain. At embryonic day 15, SLCO1C1 expression has been documented, indicating its early developmental importance .

What validated applications exist for SLCO1C1 antibodies in research?

SLCO1C1 antibodies have been validated for numerous research applications, including:

• Western Blot (WB): For detection and quantification of SLCO1C1 protein in tissue lysates

• Enzyme-Linked Immunosorbent Assay (ELISA): For quantitative measurement in solution

• Immunocytochemistry (ICC): For cellular localization studies

• Immunofluorescence (IF): For visualization in tissue sections with fluorescent detection

• Immunohistochemistry (IHC): For localization studies in fixed tissue sections

• Flow Cytometry (FCM): For cell sorting and quantification

• Immunoprecipitation (IP): For protein isolation and interaction studies

The selection of an appropriate application depends on the specific research question, sample type, and available equipment. Multiple antibody suppliers offer products validated for these various applications, with some antibodies specifically optimized for particular techniques.

What methodological considerations are essential when selecting SLCO1C1 antibodies?

When selecting SLCO1C1 antibodies for research, several methodological considerations are critical:

• Species reactivity: Verify the antibody's reactivity with your species of interest. Available antibodies vary in their cross-reactivity with human, mouse, rat, and other species .

• Antibody type: Consider whether polyclonal or monoclonal antibodies better suit your experiment. Polyclonal antibodies often provide higher sensitivity but potentially lower specificity compared to monoclonals.

• Epitope location: Some antibodies target specific regions of SLCO1C1, such as the N-terminal region (aa 11-274) or middle segments (aa 180-229) . This is particularly important when studying specific domains or potential splice variants.

• Conjugation status: Determine whether unconjugated antibodies or those conjugated to detection tags (such as Alexa Fluor 546) are more appropriate for your application .

• Validation data: Review available validation data, including published citations and supplier-provided data such as Western blot images or immunostaining patterns.

• Technical compatibility: Ensure compatibility with your specific protocols, including fixation methods, antigen retrieval requirements, and detection systems.

How should researchers optimize immunohistochemistry protocols for SLCO1C1 detection in brain tissue?

Optimizing immunohistochemistry for SLCO1C1 in brain tissue requires attention to several critical parameters:

• Fixation: Overfixation can mask epitopes; consider shorter fixation times (12-24 hours) with 4% paraformaldehyde or test alternative fixatives for epitope preservation.

• Antigen retrieval: As a membrane protein, SLCO1C1 often requires robust antigen retrieval. Test heat-induced epitope retrieval with citrate buffer (pH 6.0) and/or EDTA buffer (pH 9.0).

• Permeabilization: Optimize detergent concentration (typically 0.1-0.3% Triton X-100) to balance membrane protein accessibility with tissue morphology preservation.

• Blocking: Implement dual blocking with both serum (5-10%) and bovine serum albumin (1-3%) to minimize non-specific binding.

• Antibody dilution: Titrate primary antibodies to determine optimal concentration, typically starting with dilutions between 1:100 and 1:500.

• Incubation conditions: Compare overnight incubation at 4°C versus room temperature incubation for signal-to-noise optimization.

• Detection system: For fluorescent detection, select secondary antibodies with minimal cross-reactivity to brain tissue and consider signal amplification systems for low abundance detection.

• Controls: Include positive controls (tissues known to express SLCO1C1, such as choroid plexus) and negative controls (antibody omission, pre-adsorption with antigen, or ideally, tissue from SLCO1C1-knockout animals).

How can researchers distinguish between non-specific binding and true SLCO1C1 signal?

The challenge of non-specific binding is particularly relevant in SLCO1C1 research due to its expression in highly vascularized barriers. To distinguish true signal from artifacts:

• Multiple antibody validation: Use multiple antibodies targeting different epitopes of SLCO1C1 and compare staining patterns.

• Complementary techniques: Validate protein detection with mRNA localization techniques such as in situ hybridization or RNAscope.

• Knockout controls: When available, use SLCO1C1 knockout tissues as negative controls to definitively identify non-specific signals.

• Antigen competition assays: Pre-incubate the antibody with purified SLCO1C1 protein or peptide to block specific binding sites.

• Advanced microscopy: Use confocal microscopy to improve signal resolution and eliminate out-of-focus fluorescence that can contribute to background.

• Double fluorescent staining: Apply the strategy mentioned in search result , where double fluorescent staining (using two different fluorescent labels on the same target) can increase specificity, though this approach still requires validation as it may not eliminate all non-specific binding.

• Signal thresholding: Establish clear criteria for distinguishing positive from negative signals based on intensity quantification and comparison to established controls.

What are effective strategies for studying SLCO1C1 at the blood-brain barrier?

The blood-brain barrier presents unique challenges for SLCO1C1 research due to its complex cellular architecture. Effective strategies include:

• Co-localization studies: Use established BBB markers (e.g., CD31, GLUT1, claudin-5) alongside SLCO1C1 antibodies to confirm vascular localization.

• 3D reconstruction techniques: Employ confocal z-stacks with 3D reconstruction to visualize the complete vascular network and SLCO1C1 distribution.

• Fresh tissue techniques: Use vibratome sections of lightly-fixed tissue to preserve antigenicity of membrane proteins like SLCO1C1.

• In vitro BBB models: Utilize primary brain endothelial cell cultures or immortalized BBB cell lines to study SLCO1C1 function under controlled conditions.

• Transgenic reporter approaches: Consider transgenic approaches similar to the Slco1c1-Cre mice described in the search results , which allow for cell-specific labeling of SLCO1C1-expressing cells.

• Electron microscopy: For subcellular localization, immunogold labeling with SLCO1C1 antibodies can provide high-resolution information about transporter positioning relative to luminal and abluminal membranes.

• Brain region comparison: Systematically compare SLCO1C1 expression across brain regions with varying BBB properties to correlate with functional differences.

How can researchers address contradictory results from different SLCO1C1 antibodies?

When confronted with contradictory results from different antibodies, implement a systematic troubleshooting approach:

• Epitope mapping: Determine the exact epitopes recognized by each antibody and evaluate whether they might be differentially accessible under various experimental conditions.

• Technical validation matrix: Create a matrix comparing antibody performance across multiple technical parameters (fixation methods, antigen retrieval protocols, detection systems, etc.).

• Orthogonal techniques: Implement non-antibody-based methods such as RNA-seq, RT-PCR, or mass spectrometry to independently confirm SLCO1C1 expression.

• Species considerations: Verify that discrepancies aren't due to species differences, as SLCO1C1 sequence conservation varies across species.

• Post-translational modifications: Consider whether post-translational modifications might affect epitope accessibility for certain antibodies.

• Isoform specificity: Determine whether contradictions result from differential recognition of SLCO1C1 isoforms or splice variants.

• Published literature cross-reference: Compare your results with published findings using the same antibodies to identify methodological variables that might explain discrepancies.

What experimental approaches best capture developmental changes in SLCO1C1 expression?

To effectively document developmental changes in SLCO1C1 expression:

• Temporal sampling: Collect samples across multiple developmental timepoints spanning embryonic, early postnatal, juvenile, and adult stages.

• Cell-type specific analysis: Combine SLCO1C1 antibody labeling with cell-type specific markers to track expression transitions between cell populations during development.

• Quantitative analysis: Implement standardized quantification methods using image analysis software to objectively measure expression changes.

• Single-cell techniques: Consider single-cell RNA sequencing paired with protein detection to capture cell-specific expression dynamics.

• Lineage tracing: Utilize the transgenic approach mentioned in search result , where Slco1c1-Cre mice crossed with reporter mice revealed transient expression in neuronal populations during development.

• Functional correlation: Correlate SLCO1C1 expression patterns with functional readouts such as local thyroid hormone levels or activation of thyroid hormone-responsive genes.

• Regional analysis: Develop brain region-specific analyses to capture spatial heterogeneity in developmental expression patterns.

Expression levels: - absent, + low, ++ moderate, +++ high
(Table derived from data in search result and developmental expression patterns)

How can SLCO1C1 antibodies be integrated with advanced molecular techniques?

Modern molecular biology offers numerous opportunities to enhance SLCO1C1 antibody research:

• Proximity ligation assays: Use antibody-based proximity ligation to detect and visualize SLCO1C1 interactions with binding partners or regulatory proteins.

• CRISPR/Cas9 epitope tagging: Generate endogenously tagged SLCO1C1 to enable detection without relying solely on antibody specificity.

• ChIP-seq integration: Combine chromatin immunoprecipitation with SLCO1C1 expression analysis to correlate transcriptional regulation with protein expression.

• Protein-fragment complementation: Develop split-reporter systems fused to SLCO1C1 to study dynamic protein interactions in live cells.

• Fluorescence correlation spectroscopy: Apply FCS with fluorescently-labeled SLCO1C1 antibodies to study transporter dynamics and mobility.

• Expansion microscopy: Physically expand specimens to improve resolution of SLCO1C1 localization in complex brain structures.

• Deep learning image analysis: Implement machine learning approaches to extract complex patterns from SLCO1C1 immunostaining data.

What considerations are important when using SLCO1C1 antibodies in human pathological specimens?

Working with human pathological specimens introduces additional methodological challenges:

• Post-mortem interval effects: Validate SLCO1C1 antibody performance across different post-mortem intervals to establish detection reliability.

• Fixation variability: Optimize protocols to accommodate variations in fixation methods and durations common in clinical specimens.

• Disease-specific changes: Consider how pathological conditions might alter SLCO1C1 epitope accessibility or expression.

• Batch normalization: Implement rigorous normalization procedures when comparing specimens collected under variable conditions.

• Age-matched controls: Ensure appropriate age-matching of control specimens due to age-related changes in SLCO1C1 expression.

• Regional sampling strategy: Develop consistent anatomical sampling strategies to account for region-specific expression patterns.

• Clinical correlation: Design studies that correlate SLCO1C1 immunohistochemical findings with clinical parameters and outcomes.

What are common pitfalls in SLCO1C1 antibody validation and how can they be avoided?

Antibody validation is crucial for reliable SLCO1C1 research. Common pitfalls and solutions include:

• Insufficient specificity testing: Implement comprehensive controls including knockout/knockdown tissues, peptide competition, and multiple antibodies targeting different epitopes.

• Inadequate positive controls: Identify and include tissues with established SLCO1C1 expression (e.g., choroid plexus) as positive controls in all experiments.

• Non-optimized protocols: Systematically optimize fixation, antigen retrieval, and detection parameters for each new antibody rather than applying standardized protocols.

• Cross-reactivity with related proteins: Test for cross-reactivity with other SLCO family members, particularly in tissues where multiple transporters are expressed.

• Batch-to-batch variability: Document antibody lot numbers and validate new lots against previously characterized lots.

• Limited application validation: Validate each antibody specifically for your intended application rather than assuming cross-application reliability.

• Inadequate reporting: Maintain detailed records of validation procedures and results to ensure reproducibility and transparency.

How should researchers interpret discrepancies between protein detection (antibody-based) and transcript detection of SLCO1C1?

Discrepancies between protein and mRNA detection are common and provide important biological insights:

• Post-transcriptional regulation: Consider whether microRNAs or RNA-binding proteins might regulate SLCO1C1 translation efficiency.

• Protein stability differences: Evaluate protein half-life as a factor in accumulation or depletion relative to transcript levels.

• Temporal dynamics: Assess whether sampling timing captures transient expression differences between mRNA and protein.

• Cellular compartmentalization: Verify whether detection methods adequately capture protein in all cellular compartments, particularly for membrane proteins like SLCO1C1.

• Technical sensitivity differences: Compare detection thresholds between antibody-based and nucleic acid-based methods.

• Cell-type heterogeneity: Consider whether bulk tissue analysis masks cell-type specific differences in translation efficiency.

• Experimental validation: Design experiments that specifically track both transcript and protein levels in the same samples over time to establish correlation patterns.

What quality control measures should be implemented when using SLCO1C1 antibodies in research?

Rigorous quality control is essential for SLCO1C1 antibody research:

Comparison of SLCO1C1 Antibody Applications in Research Settings