SLC52A3 Antibody
Description
Definition and Purpose of SLC52A3 Antibody
The SLC52A3 antibody is a polyclonal or monoclonal reagent designed to bind specifically to the SLC52A3 protein (solute carrier family 52, riboflavin transporter, member 3). This protein is encoded by the SLC52A3 gene and facilitates riboflavin absorption in the small intestine, enabling its conversion into flavin coenzymes (FAD/FMN) for energy metabolism . Antibodies targeting SLC52A3 are primarily used in research to:
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Localize the protein in tissues (e.g., small intestine, brain, cancer cells)
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Investigate diseases linked to SLC52A3 mutations, such as Brown-Vialetto-Van Laere syndrome (BVVL)
Applications in Research
SLC52A3 antibodies are validated for multiple experimental techniques:
Role in Neurological Disorders
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BVVL Syndrome: SLC52A3 antibodies helped identify reduced riboflavin transport in patients with SLC52A3 mutations, leading to mitochondrial dysfunction and neurodegeneration . Autopsy studies revealed brainstem neuron loss in affected individuals .
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Mouse Models: Slc52a3 knockout mice exhibited neonatal lethality, cortical hypoplasia, and riboflavin deficiency, detected via antibody-based tissue analysis .
Cancer Research
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Esophageal Squamous Cell Carcinoma (ESCC): SLC52A3 antibodies confirmed overexpression of the protein in ESCC tissues. High SLC52A3 levels correlated with poor prognosis and NF-κB-driven tumor proliferation .
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Therapeutic Target: Riboflavin supplementation reversed symptoms in BVVL patients, highlighting the antibody’s utility in monitoring treatment efficacy .
Validation and Quality Control
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
The SLC52A3 polyclonal antibody is produced by immunizing a rabbit with a synthetic peptide encompassing amino acids 159-220 of the recombinant human SLC52A3 protein. The resulting polyclonal antibody mixture undergoes purification via protein G affinity chromatography, achieving a purity exceeding 95%. The purified SLC52A3 antibody undergoes stringent testing for specificity and affinity through ELISA, Western Blot, Immunohistochemistry, and Immunofluorescence assays to ensure its capacity to bind to the SLC52A3 protein. This antibody demonstrates reactivity with both human and mouse samples.
The target protein SLC52A3 primarily functions as a high-affinity riboflavin transporter, facilitating the uptake of riboflavin from the extracellular environment into cells. This process is critical for maintaining adequate intracellular and tissue riboflavin levels. Notably, SLC52A3 has also been implicated in the regulation of cell growth and proliferation within cancer cells.
Target Background
- The 5'-flanking regions of the riboflavin transporter-3 (SLC52A3) gene contain NF-kappaB p65/Rel-B-binding sites, which are essential for mediating SLC52A3 transcriptional activity in esophageal squamous cell carcinoma (ESCC) cells. PMID: 29428966
- Gene and protein expression levels of RFVT3 were found to be higher in DLD-1 and HT-29 cell lines compared to Caco2 cells. In tumor tissues of patients diagnosed with colorectal cancer (CRC), RFVT3 gene expression levels were elevated, while protein expression was reduced, accompanied by a slight decrease in riboflavin content. PMID: 29715086
- This study presents the first evidence suggesting that Rfvt3 contributes to riboflavin transport in the placenta, and that disruption of the Slc52a3 gene results in neonatal mortality associated with hyperlipidemia and hypoglycemia due to riboflavin deficiency. PMID: 27272163
- This research uncovered that RFT2 was overexpressed in glioma samples compared to normal brain tissue. PMID: 27584688
- RFVT3 is a target for posttranscriptional regulation by miR-423-5p in intestinal epithelial cells, and this regulation has functional consequences on intestinal riboflavin (RF) uptake. PMID: 28912250
- Fourteen mutations in SLC52a3 were linked to Brown-Vialetto-Van Laere syndrome. PMID: 29053833
- The SLC52A3 rs13042395 C>T polymorphism was associated with a reduced cancer risk in individuals with a normal body mass index, while no association was observed in the obese group. PMID: 27600099
- Carriers of the SLC52A3 rs13042395 TT genotype demonstrated a reduced likelihood of lymph node metastasis and a longer relapse-free survival time. PMID: 27472962
- This study identified several residues within the hRFVT-3 polypeptide that are crucial for its functionality and cell surface expression. PMID: 28637675
- RFT2 plays a significant role in gastric carcinogenesis by modulating riboflavin absorption. PMID: 26722538
- The C20orf54 rs13042395 polymorphism was significantly associated with a decreased risk of ESCC and GCA, particularly among underweight or normal weight individuals. PMID: 26154995
- A strong association exists between the functional SNP rs3746804 in C20orf54 and susceptibility to esophageal squamous cell carcinoma. PMID: 25427582
- The single-nucleotide polymorphism rs13042395 in C20orf54 exhibited a significantly lower risk of esophageal squamous cell carcinoma in the younger age group, but no significant association was observed in the older group within a Korean population. PMID: 24152165
- Binding of Sp1 to the minimal SLC52A3 promoter has been observed. PMID: 25394472
- Increased methylation of CpG 2 and CpG 3 in the hRFT2 gene promoter region is associated with the development of cervical squamous cell carcinoma. PMID: 24761851
- Research suggests that RFT2 contributes to esophageal squamous cell carcinoma tumorigenesis and may serve as a potential therapeutic target. PMID: 25045844
- C20orf54 expression levels were significantly upregulated in CSCC. PMID: 24260322
- These findings strongly suggest that RFVT3 plays a functional role in riboflavin absorption in the apical membranes of intestinal epithelial cells. PMID: 24264046
- Defective expression of C20orf54 is associated with the development of Kazak esophageal squamous cell carcinoma, which may represent a mechanism underlying the decreased plasma riboflavin levels observed in ESCC. PMID: 23275236
- The functional single nucleotide polymorphism in the RFT2 protein might be associated with the development of esophageal squamous cell carcinoma. PMID: 22533825
- Identification of novel mutations that affect amino acid changes in patients with Brown-Vialetto-Van Laere syndrome has been reported. PMID: 22718020
- Defective expression of RFT2 is associated with the development of gastric carcinoma, potentially leading to decreased plasma riboflavin levels in GC. PMID: 22791947
- Single nucleotide polymorphism in the C20orf54 gene has been linked to esophageal squamous cell carcinoma. PMID: 22471455
- Mutations in the riboflavin transporter-2 gene have been associated with Brown-Vialetto-Van Laere syndrome. PMID: 22273710
- Research has demonstrated a potential role for specific cysteine residues in the cell surface expression of riboflavin transporter 2 in human intestinal epithelial cells. PMID: 21512156
- Findings indicate that riboflavin transporter 2 (RFT2) is a transporter involved in the epithelial uptake of riboflavin in the small intestine, facilitating its nutritional utilization. PMID: 20724488
- Susceptibility loci at C20orf54 have been identified for esophageal squamous cell carcinoma. PMID: 20729853
- This study discusses the cloning of rat riboflavin transporter 2 and the identification of a comparable protein in humans. PMID: 19122205
- A candidate gene, C20orf54, was identified in a consanguineous family with Brown-Vialetto-Van Laere syndrome, involving multiple affected individuals. Subsequent investigations demonstrated that mutations in this gene were the causative factor for the disease in other, unrelated families. PMID: 20206331
Q&A
What are the primary applications of SLC52A3 antibodies in immunodetection?
SLC52A3 antibodies are primarily used for:
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Western Blot (WB): Detecting the 51 kDa protein in lysates from tissues with high expression (e.g., testis, intestine, placenta) .
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Immunohistochemistry (IHC): Localizing SLC52A3 in paraffin-embedded tissues (e.g., human stomach/esophagus cancer, mouse testis) using antigen retrieval with TE buffer (pH 9.0) or citrate buffer (pH 6.0) .
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ELISA: Quantifying SLC52A3 levels in biological fluids, though validation data remains limited .
| Application | Recommended Dilution | Key Considerations |
|---|---|---|
| WB | 1:500–1:1000 | Use PVDF membranes; optimize protein loading (30–50 µg) |
| IHC | 1:500–1:2000 | Perform antigen retrieval; validate with adjacent sections |
How do I optimize antibody dilution for SLC52A3 detection?
Optimal dilution requires iterative testing:
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WB: Start with 1:500 (0.2 µg/mL) and titrate upward if signal is weak .
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IHC: Begin with 1:500 (0.4 µg/mL) and increase to 1:2000 (0.1 µg/mL) for stronger signals in thick tissues .
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Controls: Include SLC52A3-positive lysates (e.g., mouse brain tissue) and negative controls (e.g., SLC52A3 knockout samples) .
What factors influence cross-reactivity of SLC52A3 antibodies?
Cross-reactivity depends on:
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Immunogen Design: Antibodies targeting C-terminal regions (e.g., AA 391–469) show stronger cross-reactivity with mouse/rat .
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Host Species: Rabbit polyclonal antibodies exhibit broader reactivity (human, mouse, rat) compared to monoclonals .
| Antibody | Tested Reactivity | Predicted Reactivity |
|---|---|---|
| ABIN754768 | Mouse, Rat | Human, Dog, Cow |
| 25626-1-AP | Human, Mouse | — |
How can SLC52A3 antibodies be used to study developmental biology?
SLC52A3 antibodies are critical for analyzing early embryogenesis:
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Placental Development: Stain E10.5 mouse embryos to assess SLC52A3 expression in extraembryonic tissues (e.g., placenta, chorion) .
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Apoptosis Analysis: Combine TUNEL staining with SLC52A3 IHC to link riboflavin transport deficits to programmed cell death in Slc52a3−/− mutants .
What validation strategies ensure SLC52A3 antibody specificity?
Validation protocols include:
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Western Blot: Use SLC52A3 fusion protein (Ag22315) as a positive control and Slc52a3 knockout lysates as negatives .
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IHC: Perform peptide competition assays (e.g., pre-incubate antibody with immunizing peptide) to eliminate nonspecific binding .
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Bioinformatics: Align antibody epitopes with conserved regions of SLC52A3 orthologs to predict cross-reactivity .
How do I resolve contradictions in SLC52A3 antibody performance?
Common discrepancies and solutions:
What advanced applications exist for SLC52A3 antibodies in disease modeling?
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Brown-Vialetto-Van Laere Syndrome (BVVL): Use antibodies to assess riboflavin uptake in patient-derived fibroblasts or induced pluripotent stem cells (iPSCs) .
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Cancer Research: Investigate SLC52A3 overexpression in gastrointestinal cancers (e.g., stomach/esophagus) via IHC .
How do SLC52A3 antibodies compare to other riboflavin transporters (SLC52A1/A2)?
What troubleshooting steps address non-specific binding in SLC52A3 IHC?
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Blocking Agents: Use 5% BSA instead of serum to reduce background in human cancer tissues .
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Antigen Retrieval: For paraffin sections, validate with both TE (pH 9.0) and citrate (pH 6.0) buffers .
