SLC13A3 Antibody
Description
Comprehensive Overview of SLC13A3 Antibody: Characteristics, Applications, and Research Findings
The SLC13A3 antibody represents a crucial investigative tool in biochemical and biomedical research, enabling scientists to detect, visualize, and quantify the SLC13A3 protein—a sodium-dependent dicarboxylate transporter with significant roles in cellular metabolism. These antibodies have been instrumental in advancing our understanding of SLC13A3's functions in normal physiology and its implications in various pathological conditions, particularly liver cancers with specific genetic mutations. Recent research has positioned SLC13A3 not only as a marker for certain disease states but also as a potential therapeutic target, highlighting the growing importance of SLC13A3 antibodies in both basic science and translational medicine applications.
Definition and Basic Characteristics
The SLC13A3 antibody is an immunoglobulin specifically designed to recognize and bind to the SLC13A3 protein, a member of the solute carrier family 13. SLC13A3, also known as NaC3, NADC3, or SDCT2, functions as a sodium-dependent dicarboxylate cotransporter located primarily in the cell membrane . These antibodies are typically developed in host animals such as rabbits and are available in various forms, including polyclonal variants that recognize multiple epitopes of the target protein .
SLC13A3 antibodies serve as essential reagents in laboratory research, enabling the detection and analysis of SLC13A3 expression across different cell types and tissues. The specificity of these antibodies allows researchers to investigate the protein's involvement in critical cellular processes, particularly those related to metabolism and transport functions .
Target Protein: SLC13A3
SLC13A3 encodes the plasma membrane Na+/Dicarboxylate Cotransporter 3 (NaDC3), which imports four to six carbon dicarboxylates as well as N-acetylaspartate (NAA) into cells . This protein plays a crucial role in transporting Krebs cycle intermediates such as citrate and succinate, thereby influencing multiple metabolic pathways including fatty acid synthesis, glucose utilization, and cellular energy production .
While SLC13A3 is predominantly expressed in kidney tissues, it also appears in brain, liver, placenta, and eye tissues . The protein's involvement in various physiological processes makes it a significant target for research into metabolic disorders and other pathological conditions. Recent studies have particularly highlighted its role in liver cancer pathogenesis, suggesting potential therapeutic applications for targeting this protein .
Validated Research Applications
SLC13A3 antibodies have been validated for multiple laboratory applications, allowing researchers to study this protein through various experimental approaches. Primary applications include Western blotting (WB), immunohistochemistry (IHC), and enzyme-linked immunosorbent assay (ELISA) .
In Western blot applications, these antibodies enable researchers to detect and semi-quantify SLC13A3 protein levels in cell and tissue lysates. For immunohistochemistry, they allow visualization of SLC13A3 distribution in tissue sections, providing insights into its localization and expression patterns in different physiological and pathological states .
Recommended Protocols and Dilutions
Proper antibody dilutions are critical for obtaining specific and reproducible results across different applications. Table 2 presents the recommended dilutions for SLC13A3 antibodies in various applications.
Table 2: Recommended Dilutions for SLC13A3 Antibodies
| Application | SLC13A3 Antibody (CAB15183) | SLC13A3 Antibody (26182-1-AP) |
|---|---|---|
| Western Blot (WB) | 1:500 - 1:2000 | 1:500 - 1:2000 |
| Immunohistochemistry (IHC) | Not specified | 1:50 - 1:500 |
| ELISA | Not specified | Not specified |
For optimal immunohistochemistry results with the 26182-1-AP antibody, antigen retrieval with TE buffer pH 9.0 is suggested, with citrate buffer pH 6.0 as an alternative . The positive samples validated for Western blot applications include U-251MG and 293T cells for CAB15183 , and mouse kidney tissue and HEK-293 cells for 26182-1-AP .
Detailed protocols for these applications are typically provided by the manufacturers, with specific optimization recommendations for each experimental context to achieve optimal signal-to-noise ratio and specific detection .
Role in Normal Physiology
SLC13A3 plays essential roles in cellular metabolism through its function as a sodium-dependent dicarboxylate transporter. The protein is primarily responsible for importing Krebs cycle intermediates such as succinate and citrate into cells, contributing to various metabolic pathways including fatty acid synthesis, glucose utilization, and energy production .
Functional studies using SLC13A3-overexpressing HEK293 cells have demonstrated a significant increase in the uptake of 14C-succinate compared to control cells, confirming the transporter activity of this protein . Metabolomic profiling of these cells has revealed altered metabolite profiles, highlighting the impact of SLC13A3 on cellular metabolism .
The expression pattern of SLC13A3 across tissues—predominantly in kidney but also in brain, liver, placenta, and eye—suggests tissue-specific functions of this transporter in different physiological contexts . The ability to detect and study these expression patterns using SLC13A3 antibodies has been instrumental in elucidating these functions.
Implications in Cancer Research
Recent research has uncovered significant correlations between SLC13A3 expression and cancer, particularly hepatocellular carcinoma (HCC). Analysis of multiple human HCC gene expression datasets has revealed that SLC13A3 mRNA expression is upregulated in cirrhotic tissues and further increased in HCC tissues compared to healthy liver tissues .
Co-expression network analysis has demonstrated substantial interactions between SLC13A3 and several canonical β-catenin target genes, such as LGR5, TBX3, and GLUL, suggesting a potential relationship with the Wnt/β-catenin signaling pathway . This connection is further supported by the observation that HCCs with high SLC13A3 expression are mainly characterized by CTNNB1 mutations .
Immunohistochemistry studies utilizing SLC13A3 antibodies have shown that SLC13A3 protein levels are higher in HCCs with cytoplasmic/nuclear β-catenin expression compared to their paired normal counterparts . This cytoplasmic/nuclear accumulation of β-catenin, corresponding to hyperactivation of Wnt/β-catenin signaling, is significantly associated with SLC13A3 overexpression .
Molecular Mechanisms and Pathways
SLC13A3 knockdown experiments have provided insights into the molecular mechanisms through which this protein influences cellular processes. These studies have revealed that SLC13A3 knockdown significantly decreases glutathione (GSH) levels, an important cellular antioxidant .
Furthermore, SLC13A3 knockdown has been shown to affect DNA methylation, with a marked increase in 5-methylated-cytosine (5-mC) in the MYC promoter . Methylation-specific PCR analysis has revealed that methylation of CpG islands in the MYC promoter region increases in SLC13A3-knockdown cells and decreases in SLC13A3-overexpressing cells, suggesting an epigenetic mechanism through which SLC13A3 regulates gene expression .
Additionally, SLC13A3 appears to influence autophagy and ferroptosis, a form of regulated cell death. SLC13A3 knockdown has been observed to decrease ferritin heavy chain 1 (FTH1) and increase ATG5, suggesting the occurrence of ferritinophagy . These findings highlight the complex involvement of SLC13A3 in multiple cellular pathways and processes.
Diagnostic Potential
The correlation between SLC13A3 expression and specific cancer mutations presents potential diagnostic applications for SLC13A3 antibodies. Studies have shown that SLC13A3 expression is significantly higher in CTNNB1 mutant HCCs compared to non-CTNNB1 mutant HCCs, suggesting that SLC13A3 could serve as a biomarker for this genetic subtype of liver cancer .
Therapeutic Implications
Research findings suggesting that SLC13A3 is a major effector downstream of activated β-catenin in liver cancer point to its potential as a therapeutic target . The relationship between SLC13A3 expression and various metabolic and cellular processes further supports this potential.
Experimental evidence from SLC13A3 knockdown studies has demonstrated decreased tumor cell proliferation and increased ferroptosis, suggesting that targeting SLC13A3 could have anti-tumor effects . These studies highlight the therapeutic potential of strategies aimed at modulating SLC13A3 function or expression in specific cancers, particularly those characterized by CTNNB1 mutations .
Product Specs
Target Background
This antibody targets NaDC3 (SLC13A3), a high-affinity sodium-dicarboxylate cotransporter. It exhibits broad substrate specificity, transporting molecules with 4-6 carbon atoms, including succinate, α-ketoglutarate, and N-acetylaspartate. The likely stoichiometry is 3 Na+ ions per 1 divalent succinate molecule.
- Immunostaining reveals NaDC3 localization in the basolateral membrane of proximal tubules, and in both the basolateral and luminal membranes of principal cells within connecting segments and collecting ducts. (PMID: 27053689)
- OAT1, NaDC3 (basolateral membrane), and OAT4 (luminal membrane) in proximal tubule cells mediate the renal secretion of N-carbamoylglutamate. (PMID: 25354943)
- A study on chronic kidney disease in Sri Lanka identified SNP rs6066043 within the SLC13A3 gene as having a 50% attributable risk. (PMID: 24351856)
- NaDC3, located on the basolateral membrane of proximal tubule cells, mediates sodium-dependent glutathione (GSH) uptake, functioning as a low-affinity GSH transporter. (PMID: 24247155)
- α-ketoglutarate is a high-affinity substrate shared by NaDC3, OAT1, and OAT3. (PMID: 21865262)
- SLC13A3 is a direct downstream target of PITX2 transcriptional regulation, and their levels modulate cellular responses to oxidative stress in ocular cells. (PMID: 21873665)
- NaDC3 may promote cellular senescence by inhibiting NAD+-dependent SIRT1. (PMID: 20813124)
- The specific substrate selectivity of NaDC3 prevents drug interactions with dicarboxylate-like structures, ensuring adequate α-ketoglutarate supply to proximal tubule cells for anion secretion via OAT1 or OAT3. (PMID: 15561973)
- NaDC3 is localized to the basolateral membrane of human renal proximal tubule cells; a di-hydrophobic amino acid motif (VW) in its N-terminal cytoplasmic domain serves as a basolateral localization signal. (PMID: 16331647)
Q&A
Basic Research Questions about SLC13A3 Antibody
What are the optimal conditions for using SLC13A3 antibodies in immunohistochemistry?
For optimal IHC results with SLC13A3 antibodies:
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Antigen retrieval: Use TE buffer at pH 9.0 as the primary method; citrate buffer at pH 6.0 can be used as an alternative
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Antibody dilution: Start with 1:200-1:500 for polyclonal antibodies and optimize based on signal strength and background
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Positive control tissues: Mouse or human kidney tissue sections are recommended
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Detection systems: Both DAB (3,3'-diaminobenzidine) and fluorescent secondary antibodies have been validated
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Incubation conditions: Overnight incubation at 4°C often yields optimal results for membrane proteins
When analyzing liver cancer samples, researchers should note the correlation between SLC13A3 expression and β-catenin localization; cytoplasmic/nuclear accumulation of β-catenin significantly associates with SLC13A3 overexpression (R = 0.9084, P < 0.0001) .
How can I validate the specificity of my SLC13A3 antibody?
Comprehensive validation of SLC13A3 antibodies should include:
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Positive controls:
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Negative controls:
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Genetic validation approaches:
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Functional validation:
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Cross-reactivity assessment:
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Test against other SLC13A family members (SLC13A1-5) to ensure specificity
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Researchers should observe the expected molecular weight (55-70 kDa depending on the system and glycosylation state) .
How can SLC13A3 antibodies be used to study the Wnt/β-catenin pathway in liver cancer?
SLC13A3 antibodies provide valuable tools for investigating Wnt/β-catenin pathway activation in liver cancer through several methodological approaches:
Why might I observe different molecular weights for SLC13A3 in different tissue samples?
Variation in observed molecular weights for SLC13A3 across different samples is common and may result from several biological and technical factors:
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Post-translational modifications:
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Glycosylation patterns differ between tissues and cell types
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Phosphorylation states can alter migration patterns
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Ubiquitination or SUMOylation may create higher molecular weight bands
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Tissue-specific isoforms:
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Alternative splicing can generate tissue-specific variants
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Different promoter usage may result in truncated protein forms
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Technical considerations:
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Sample preparation methods (reducing vs. non-reducing conditions)
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Gel percentage and running conditions
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Protein loading amount and saturation effects
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The expected molecular weight ranges documented in the literature:
To address these variations:
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Always include appropriate positive controls
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Consider using multiple antibodies targeting different epitopes
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Perform validation with genetic manipulation (overexpression/knockdown)
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Include tissue-specific controls when comparing across different sample types
How can I differentiate between SLC13A3 and other SLC13A family members in my experiments?
Distinguishing SLC13A3 from other SLC13A family members (SLC13A1-5) requires careful experimental design:
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Antibody selection strategies:
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Choose antibodies raised against unique regions with minimal sequence homology
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Target the N- or C-terminal regions which tend to be less conserved
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Validate specificity using overexpression systems for each family member
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Molecular techniques for validation:
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Functional differentiation:
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Expression pattern analysis:
When analyzing RNA-seq or microarray data, researchers should verify that probes or sequencing reads map uniquely to SLC13A3 rather than conserved regions shared with other family members.
What are common pitfalls when interpreting SLC13A3 expression data in cancer studies?
Researchers analyzing SLC13A3 expression in cancer contexts should be aware of several interpretative challenges:
To address these challenges, researchers should:
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Stratify samples by molecular subtype before comparing expression
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Use multiple detection methods (IHC, WB, qPCR)
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Include functional assays alongside expression analysis
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Consider metabolic context when interpreting transporter activity
How can SLC13A3 antibodies be utilized in therapeutic development research?
SLC13A3 antibodies serve as essential tools in developing and evaluating potential therapeutic strategies:
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Target validation studies:
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Demonstrate expression in disease-relevant tissues
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Visualize subcellular localization to confirm accessibility
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Quantify expression levels in patient samples to stratify potential responders
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Pharmacodynamic marker development:
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Therapeutic antibody development:
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Identify accessible epitopes for potential therapeutic antibody generation
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Screen for antibodies that inhibit transport function
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Evaluate internalization kinetics of antibody-SLC13A3 complexes
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Combination therapy assessment:
Research indicates that targeting SLC13A3 could be particularly effective in two cancer contexts:
