SLC6A12 Antibody

What is SLC6A12 and why is it important in neuroscience research?

SLC6A12 (Solute Carrier Family 6 Member 12) is a transporter that mediates the cellular uptake of betaine and GABA in a sodium- and chloride-dependent process. It plays a critical role in the regulation of GABAergic transmission in the brain through the reuptake of GABA into presynaptic terminals and is also involved in osmotic regulation in renal and hepatic tissues . Recent research has revealed that SLC6A12 is highly enriched in brain pericytes, making it an important marker for these cells in neurovascular research . This unexpected localization provides new opportunities for studying blood-brain barrier function and neurovascular unit integrity in both normal physiology and pathological conditions.

What types of SLC6A12 antibodies are available for research purposes?

Several types of SLC6A12 antibodies have been developed for research applications:

• Polyclonal antibodies targeting different epitopes:

  • N-terminal antibodies (AA 22-35, AA 9-38, AA 16-46)

  • C-terminal antibodies

  • Mid-region antibodies (AA 500-600)

• Recombinant monoclonal antibody pairs designed for quantitative assays

• Species-specific antibodies with varying reactivity (human, mouse, rat)

The choice of antibody depends on the specific research application, target species, and epitope accessibility in the experimental system. For pericyte labeling in human FFPE brain sections, specific SLC6A12 antibodies have been validated and optimized .

How can SLC6A12 antibodies be used for identifying pericytes in human brain tissue?

Recent research has established SLC6A12 as a highly specific marker for brain pericytes. When using SLC6A12 antibodies for pericyte identification:

• Optimize immunohistochemical techniques specifically for FFPE human brain sections

• Use appropriate antibody concentrations (approximately 0.2 μg/ml has been validated)

• Compare staining patterns with traditional pericyte markers like PDGFRB

• Validate results with complementary approaches (e.g., immunoblots)

Studies have demonstrated that SLC6A12 antibody staining is strongly positive in small blood vessels and is more effective than PDGFRB antibody at identifying pericyte-like cells in FFPE human brain sections. Importantly, this pericyte-specific staining pattern appears to be unique to brain tissue, as exploratory samples from other human organs (kidney, lung, liver, muscle) did not show the same pericyte-like staining pattern . This tissue specificity makes SLC6A12 a valuable tool for investigating the neurovascular unit in health and disease.

What are the considerations for using SLC6A12 antibodies in cross-species studies?

When designing experiments involving multiple species, researchers must carefully select SLC6A12 antibodies with appropriate cross-reactivity profiles:

• Some antibodies are specifically designed for human samples and not recommended for other species

• Other antibodies show reactivity with human, mouse, and rat samples

• Specific epitope recognition may vary between species due to sequence differences

For example, the antibody described in source is specifically designed to recognize BGT-1 from mouse and rat samples but is unlikely to recognize the protein from human samples. Conversely, other commercially available antibodies show good reactivity with human samples but might not recognize the protein in rodent tissues with equal efficiency . When conducting cross-species studies, validation experiments should be performed for each species of interest to confirm antibody specificity and optimal working conditions.

How do SLC6A12 antibodies contribute to understanding neurovascular biology?

The recent identification of SLC6A12 as a highly specific marker for brain pericytes represents a significant advance in neurovascular research. Using optimized SLC6A12 antibodies allows researchers to:

• More accurately identify and quantify pericytes in human brain tissue

• Investigate pericyte distribution across different brain regions

• Examine pericyte alterations in neurological disorders

• Study blood-brain barrier integrity in relation to pericyte function

This approach complements single-cell RNA sequencing data that identified SLC6A12 as having highly pericyte-enriched expression. The combination of transcriptomic data with protein-level validation through antibody-based techniques provides a powerful approach for investigating the cellular composition and function of the neurovascular unit. This is particularly valuable for studying conditions where blood-brain barrier dysfunction may play a role, such as vascular cognitive impairment and dementia (VCID) .

What protocol optimizations are necessary for SLC6A12 antibody use in Western blotting?

For successful Western blot applications using SLC6A12 antibodies, consider the following protocol optimizations:

• Sample preparation:

  • Use appropriate lysis buffers that preserve membrane protein integrity

  • Include protease inhibitors to prevent degradation

  • Control protein loading (10-30 μg total protein recommended)

• Gel electrophoresis:

  • Use gradient gels (4-12% or 4-20%) for optimal separation

  • Expected molecular weight for SLC6A12/BGT-1: approximately 69 kDa

• Transfer and detection:

  • Optimize transfer conditions for membrane proteins

  • Blocking: 5% non-fat milk or BSA in PBST (PBS with 0.05% Tween-20)

  • Primary antibody dilution: 1:1000-1:4000 (optimize based on specific antibody)

  • Secondary antibody: HRP-linked anti-rabbit (1:5000-1:10000)

  • Detection systems: ECL Western Blotting Substrate or SuperSignal West Pico PLUS

Validated cell lines for positive controls include L02 cells, HSC-T6 cells, HuH-7 cells, HepG2 cells, and mouse hepatocytes . Expected band sizes should be verified against the predicted molecular weight of SLC6A12 (approximately 69 kDa), and multiple bands may indicate post-translational modifications or proteolytic processing.

How should immunohistochemistry protocols be optimized for SLC6A12 detection in brain tissue?

For optimal detection of SLC6A12 in brain tissue sections, especially for pericyte identification:

• Tissue preparation:

  • Formalin-fixed paraffin-embedded (FFPE) human brain sections work well

  • Consider antigen retrieval methods to enhance signal

• Antibody selection and optimization:

  • Select antibodies validated for IHC applications

  • Anti-SLC6A12 antibody (Atlas Antibodies, HPA034973) has been validated at 1:2000 dilution (0.2 μg/ml)

  • Perform titration experiments to determine optimal concentration

• Signal detection and validation:

  • Use appropriate secondary antibodies and detection systems

  • Include positive and negative controls

  • Compare staining patterns with known pericyte markers (e.g., PDGFRB)

  • Consider counterstaining to visualize tissue architecture

• Data interpretation:

  • SLC6A12 staining should be strongly positive in small blood vessels

  • Pericyte-like staining pattern should be distinct from endothelial or smooth muscle cells

  • This pattern is specific to brain tissue and not observed in other organs

When optimizing immunohistochemistry protocols, it's important to balance sensitivity and specificity. Background staining should be minimized while maintaining strong signal in pericytes. Comparative analysis with traditional pericyte markers can help validate the specificity of SLC6A12 staining.

What approaches can be used to validate SLC6A12 antibody specificity?

Comprehensive validation of SLC6A12 antibody specificity should include multiple complementary approaches:

• Western blot validation:

  • Confirm single band at expected molecular weight (69 kDa)

  • Include positive and negative control samples

  • Consider knockdown/knockout controls if available

• Immunohistochemical validation:

  • Compare staining patterns with known marker distribution

  • Conduct peptide competition assays

  • Perform parallel staining with independent antibodies targeting different epitopes

• Transcriptomic correlation:

  • Compare antibody staining with single-cell RNA-seq data

  • Verify that protein expression matches transcript expression patterns

• Cross-species validation:

  • Test antibody performance across relevant species

  • Consider sequence homology at the epitope level

A comprehensive validation approach combining these methods provides strong evidence for antibody specificity. For example, studies have validated SLC6A12 antibodies through a combination of single-cell RNA-seq analyses showing enriched expression in pericytes, immunoblots confirming appropriate band size, and immunohistochemical studies demonstrating the expected cellular distribution pattern .

How can researchers address common technical challenges when using SLC6A12 antibodies?

When working with SLC6A12 antibodies, researchers may encounter several technical challenges:

It's important to note that some SLC6A12 antibodies have specific species limitations. For example, certain antibodies are unlikely to recognize human SLC6A12 despite working well with mouse and rat samples . Always verify the species reactivity profile before designing experiments.

How should researchers interpret conflicting SLC6A12 antibody data between different experimental approaches?

When faced with discrepancies between different experimental approaches using SLC6A12 antibodies:

• Evaluate antibody characteristics:

  • Different antibodies may target different epitopes

  • Consider accessibility of epitopes in various experimental conditions

  • Review validation data for each antibody

• Consider biological variables:

  • SLC6A12 expression may vary between cell types, tissues, and species

  • Post-translational modifications might affect antibody recognition

  • Alternative splicing could lead to isoform-specific detection

• Assess technical factors:

  • Different applications have different sensitivity thresholds

  • Sample preparation methods may affect protein conformation

  • Fixation and embedding procedures can influence epitope accessibility

• Resolve discrepancies with complementary approaches:

  • Use multiple antibodies targeting different epitopes

  • Employ orthogonal techniques (e.g., RNA-seq, mass spectrometry)

  • Consider genetic approaches (knockdown/knockout) for definitive validation

Recent research has successfully addressed potential discrepancies by combining transcriptomic data (single-cell RNA-seq) with protein detection methods (immunoblots and immunohistochemistry) to provide concordant evidence for SLC6A12 localization in brain pericytes .

What considerations are important when selecting between polyclonal and recombinant SLC6A12 antibodies?

The choice between polyclonal and recombinant SLC6A12 antibodies depends on specific research requirements:

Polyclonal SLC6A12 Antibodies:

• Advantages:

  • Recognize multiple epitopes, potentially increasing detection sensitivity

  • Often work well across multiple applications

  • May be more robust to minor sample preparation variations

• Limitations:

  • Batch-to-batch variability

  • Potential for higher background

  • May have greater cross-reactivity

Recombinant SLC6A12 Antibodies:

• Advantages:

  • Consistent performance between batches

  • High specificity for target epitope

  • Well-suited for quantitative applications

  • Better for long-term reproducibility of results

• Limitations:

  • May be more sensitive to epitope masking or denaturation

  • Potentially lower sensitivity compared to polyclonal antibodies

For specific applications like cytometric bead arrays, matched recombinant antibody pairs provide superior performance with defined sensitivity ranges (e.g., 0.625-20 ng/mL) . For immunohistochemistry applications, particularly pericyte labeling in brain tissue, specific polyclonal antibodies have been well-validated . Consider the critical requirements of your experimental system, including sensitivity needs, quantification requirements, and long-term reproducibility when selecting between antibody types.

How might SLC6A12 antibodies advance understanding of neurovascular dysfunction in disease states?

The validation of SLC6A12 as a specific pericyte marker opens new avenues for investigating neurovascular pathology:

• Neurodegenerative diseases:

  • Quantify pericyte loss or dysfunction in Alzheimer's disease

  • Examine relationships between pericyte coverage and amyloid deposition

  • Investigate blood-brain barrier integrity in relation to pericyte abnormalities

• Cerebrovascular disorders:

  • Assess pericyte changes following stroke or ischemia

  • Study pericyte responses in small vessel disease

  • Evaluate microvascular remodeling in vascular cognitive impairment

• Developmental and pediatric research:

  • Investigate pericyte development in the maturing brain

  • Study neurovascular unit formation and stabilization

  • Examine pericyte abnormalities in developmental disorders

The availability of well-validated SLC6A12 antibodies provides researchers with tools to accurately identify and quantify pericytes in human brain tissue samples . This capability is particularly valuable for comparative studies between healthy and diseased tissue, potentially revealing pericyte-specific pathologies that may contribute to disease progression or represent therapeutic targets.

What emerging research applications might benefit from SLC6A12 antibody technology?

Several cutting-edge research areas could benefit from advanced SLC6A12 antibody applications:

• Spatial transcriptomics:

  • Combining SLC6A12 antibody staining with spatial transcriptomics

  • Mapping pericyte-specific gene expression in tissue context

  • Correlating pericyte locations with regional gene expression patterns

• Multi-omics approaches:

  • Integrating antibody-based pericyte identification with proteomics

  • Correlating pericyte density with metabolomic profiles

  • Connecting pericyte status to local inflammatory signatures

• Advanced imaging approaches:

  • Super-resolution microscopy of pericyte-endothelial interactions

  • Live imaging of pericyte dynamics using labeled antibody fragments

  • Correlative light and electron microscopy for ultrastructural analysis

• Therapeutic development:

  • Targeting drug delivery to pericytes using antibody-drug conjugates

  • Monitoring pericyte responses to therapeutic interventions

  • Developing biomarkers for neurovascular dysfunction

The highly specific nature of SLC6A12 expression in brain pericytes makes it an excellent target for these advanced applications . As antibody technologies continue to evolve, including the development of recombinant antibodies with enhanced properties, these research directions will become increasingly feasible and informative.