SLC37A1 Antibody

What is SLC37A1 and why is it relevant to research?

SLC37A1 (solute carrier family 37 member 1) is an inorganic phosphate and glucose-6-phosphate antiporter localized in the endoplasmic reticulum. The canonical human protein consists of 533 amino acid residues with a molecular mass of approximately 57.6 kDa . SLC37A1 is expressed across numerous tissues, with highest expression observed in pancreas, kidney, bone marrow, spleen, liver, and small intestine, as well as in fetal brain, liver, and spleen .

As a member of the Organophosphate:Pi antiporter (OPA) protein family (TC 2.A.1.4), SLC37A1 is believed to transport cytoplasmic glucose-6-phosphate into the endoplasmic reticulum lumen while translocating inorganic phosphate in the opposite direction . This functionality makes it a significant target for research into carbohydrate metabolism, cellular homeostasis, and potentially oncology applications.

How do SLC37A1 antibodies differ from other SLC37 family member antibodies?

SLC37A1 antibodies are specifically designed to target unique epitopes of the SLC37A1 protein, distinguishing it from other family members (SLC37A2, SLC37A3, and SLC37A4). When selecting an antibody, researchers should consider:

When selecting an antibody, verification of specificity is crucial to ensure it does not cross-react with other SLC37 family members, particularly given their sequence homology .

What are the recommended applications for SLC37A1 antibodies?

SLC37A1 antibodies have been validated for multiple experimental applications:

• Western Blot (WB): Most frequently used application, providing information about protein size and expression levels

• Immunohistochemistry (IHC-P): For detection in paraffin-embedded tissues, including skeletal muscle and colon cancer samples

• Immunocytochemistry/Immunofluorescence (ICC/IF): Effective for cellular localization studies, as demonstrated in MCF7 cells

• Enzyme-Linked Immunosorbent Assay (ELISA): For quantitative measurement of SLC37A1 protein levels

When designing experiments, consider the specific experimental questions, tissue or cell types of interest, and whether quantitative or qualitative data is needed.

What are the optimal conditions for Western blot detection of SLC37A1?

Successful Western blot detection of SLC37A1 requires attention to several technical factors:

• Sample preparation: Effective extraction from membranous ER structures requires appropriate detergents (e.g., RIPA buffer with 1% NP-40 or Triton X-100)

• Gel percentage: 10% SDS-PAGE gels are typically appropriate for the 57.6 kDa protein

• Protein transfer: Semi-dry or wet transfer methods with methanol-containing buffer for optimal transfer of membrane proteins

• Blocking: 5% non-fat milk or BSA in TBST (Tris-buffered saline with Tween-20)

• Antibody dilution: Typically 1:500-1:2000 for primary antibodies, though optimal dilution should be determined experimentally

• Controls: Include positive controls (tissues known to express SLC37A1 such as pancreas or kidney) and negative controls

Note that post-translational modifications, particularly glycosylation, may result in apparent molecular weights different from the predicted 57.6 kDa . In cases of discrepancy, validation using knockdown or knockout models is recommended.

How can I optimize immunohistochemical detection of SLC37A1 in tissue sections?

For optimal IHC-P detection of SLC37A1:

• Antigen retrieval: Heat-induced epitope retrieval using citrate buffer (pH 6.0) is often effective for exposing ER-associated epitopes

• Antibody dilution: Start with 1:100 dilution as used in published protocols

• Incubation conditions: Overnight at 4°C typically yields optimal signal-to-noise ratio

• Detection system: HRP-conjugated secondary antibodies with DAB substrate or fluorescence-based detection systems

• Counterstaining: Hematoxylin for contrast in brightfield microscopy

• Controls: Include tissues with known high expression (pancreas, kidney) as positive controls and isotype controls to assess non-specific binding

Researchers should note that expression patterns may vary significantly between tissues, with particular attention to pancreas, kidney, bone marrow, spleen, liver, and small intestine where SLC37A1 expression is highest .

What approaches are recommended for validating SLC37A1 antibody specificity?

Antibody validation is crucial for ensuring experimental rigor:

• Western blot analysis: Confirm a single band of appropriate molecular weight (57.6 kDa, though may vary due to glycosylation)

• Positive and negative control tissues: Compare tissues with known high expression (pancreas, kidney) against those with lower expression

• siRNA/shRNA knockdown: Demonstrate reduced signal following SLC37A1 knockdown

• Overexpression systems: Show increased signal in overexpression models

• Peptide competition assay: Pre-incubation with the immunizing peptide should abolish specific binding

• Cross-reactivity testing: Evaluate potential cross-reactivity with other SLC37 family members, particularly SLC37A2 which shares functional similarities

• Multiple antibody approach: Use antibodies targeting different epitopes to confirm findings

Validation approaches should be selected based on experimental context and available resources, with at least two independent methods recommended.

How can SLC37A1 antibodies be used to investigate its role in cancer cell metabolism?

SLC37A1 has been implicated in cancer metabolism through its association with phospholipid biosynthesis and its regulation by EGF in breast cancer cells . Researchers can employ SLC37A1 antibodies to:

• Comparative expression analysis: Evaluate SLC37A1 expression across normal tissues, primary tumors, and metastatic samples using IHC or tissue microarrays

• Signaling pathway studies: Investigate the EGFR/MAPK/Fos pathway regulation of SLC37A1 through combined phospho-specific antibodies for signaling components and SLC37A1 antibodies

• Co-localization studies: Perform dual-label immunofluorescence with markers of ER and other organelles to assess potential redistribution in cancer cells

• Functional correlations: Combine SLC37A1 immunodetection with metabolic assays measuring glucose-6-phosphate transport or phospholipid synthesis

• Therapeutic response monitoring: Evaluate changes in SLC37A1 expression following treatment with EGFR inhibitors or other targeted therapies

A significant research finding shows that EGF transactivates the SLC37A1 promoter sequence and induces SLC37A1 mRNA and protein expression through the EGFR/MAPK/Fos transduction pathway in ER-negative SkBr3 breast cancer cells, suggesting potential oncogenic roles .

What are the experimental considerations when studying SLC37A1 in relation to other SLC37 family members?

When investigating functional distinctions between SLC37 family members:

• Antibody panel selection: Choose antibodies with minimal cross-reactivity among family members

• Functional transport assays: SLC37A1 and SLC37A2 are Pi-linked glucose-6-phosphate antiporters, while SLC37A3 is not a G6P transporter

• Inhibitor studies: Unlike SLC37A4 (G6PT), SLC37A1 and SLC37A2 are not sensitive to chlorogenic acid inhibition, which can be used as a distinguishing characteristic

• Coupling with G6Pase-a: SLC37A1 and SLC37A2 cannot form functional complexes with G6Pase-a, in contrast to SLC37A4

• Reconstitution experiments: For advanced mechanistic studies, proteoliposome reconstitution from detergent-solubilized membrane extracts can be used to study transport activities

This comparative approach helps differentiate the specific roles of each family member within cellular metabolism .

How can SLC37A1 antibodies be applied to study its potential role in glycerol-3-phosphate metabolism?

Based on its sequence homology with bacterial glycerol-3-phosphate transporters (approximately 30%), SLC37A1 is hypothesized to catalyze the exchange of glycerol-3-phosphate against phosphate . To investigate this:

• Co-immunoprecipitation studies: Use SLC37A1 antibodies to identify interaction partners involved in glycerol-3-phosphate metabolism

• Metabolic flux analysis: Combine SLC37A1 detection with labeled glycerol-3-phosphate tracking to correlate expression with metabolic activities

• Lipid synthesis correlation: Investigate relationships between SLC37A1 expression and phospholipid biosynthesis rates, particularly in proliferating cells

• Subcellular fractionation: Use differential centrifugation to isolate ER fractions, followed by SLC37A1 immunodetection and transport assays

• Mutagenesis studies: Combine site-directed mutagenesis with immunodetection to identify critical residues for transport function

The connection to glycerol-3-phosphate metabolism is particularly relevant in proliferating tumor cells, where phospholipid biosynthesis is upregulated .

What are common challenges and solutions when working with SLC37A1 antibodies?

Remember that SLC37A1 is an ER-associated membrane protein, which may require specialized extraction and handling protocols for optimal detection .

How can SLC37A1 antibodies be effectively used in co-localization studies?

For successful co-localization studies investigating SLC37A1's precise subcellular distribution:

• Sample preparation:

  • For cultured cells: 4% paraformaldehyde fixation followed by 0.1-0.3% Triton X-100 permeabilization

  • For tissue sections: Standard FFPE processing with appropriate antigen retrieval

• Antibody selection:

  • Primary antibodies from different host species (e.g., rabbit anti-SLC37A1 and mouse anti-calreticulin for ER co-localization)

  • Validated antibody dilutions (typically 1:100 for immunofluorescence)

• Imaging parameters:

  • Confocal microscopy with appropriate filter sets

  • Sequential scanning to minimize bleed-through

  • Z-stack acquisition for three-dimensional analysis

• Quantitative analysis:

  • Pearson's correlation coefficient or Manders' overlap coefficient

  • Line profile analysis across cellular compartments

  • Distance-based co-localization metrics

• Controls:

  • Single antibody controls to establish bleed-through parameters

  • Co-localization with established ER markers (calreticulin, PDI, or KDEL-tagged proteins)

This approach has successfully demonstrated the ER localization of SLC37A1, consistent with its proposed function as a glucose-6-phosphate/phosphate antiporter in the ER membrane .

What considerations are important when designing functional studies that incorporate SLC37A1 antibody detection?

When designing integrated functional and expression studies:

• Temporal dynamics:

  • Consider time-course experiments to correlate SLC37A1 expression changes with functional outcomes

  • EGF stimulation studies should include early time points (1-2 hours) through later responses (24-48 hours)

• Experimental models:

  • Cell lines with validated SLC37A1 expression (e.g., MCF7, SkBr3 for breast cancer studies)

  • Consider comparative analysis across multiple cell types with varying baseline expression

• Combined techniques:

  • Complement antibody-based detection with mRNA analysis (qPCR, RNA-Seq)

  • Integrate transport assays in intact cells or reconstituted systems

  • Consider metabolomic analysis of relevant metabolites (glucose-6-phosphate, glycerol-3-phosphate)

• Pathway interventions:

  • EGFR pathway modulators (EGF stimulation, EGFR inhibitors)

  • MAPK pathway inhibitors

  • Fos transcription factor modulation

• Functional readouts:

  • Glucose-6-phosphate transport assays

  • Phospholipid synthesis measurement

  • Cell proliferation metrics

  • ER stress markers

These integrated approaches help establish causative relationships between SLC37A1 expression and cellular functions, particularly in contexts like cancer metabolism where EGF has been shown to induce SLC37A1 expression through specific signaling pathways .

How might SLC37A1 antibodies contribute to understanding glucose metabolism disorders?

While SLC37A1 does not appear to play a direct role in homeostatic regulation of blood glucose levels (unlike SLC37A4/G6PT) , its antibodies can still contribute to metabolic disorder research through:

• Comparative expression analysis: Investigating potential compensatory expression changes in SLC37A1 in glycogen storage diseases or diabetes models

• Tissue-specific roles: Exploring SLC37A1 functions in tissues with high expression like pancreas and liver under normal and pathological conditions

• ER stress responses: Examining SLC37A1 regulation during ER stress, which is implicated in metabolic disorders

• Metabolic pathway cross-talk: Investigating potential interplay between SLC37A1 and other glucose metabolism regulators

• Differential diagnosis: Using SLC37A antibody panels (SLC37A1-A4) to differentiate between glycogen storage disease subtypes

Understanding the molecular distinctions between SLC37A1 and SLC37A4 is particularly relevant, as only the latter contributes to interprandial blood glucose homeostasis .

What are the considerations for using SLC37A1 antibodies in cancer research?

Given the evidence for EGF-mediated upregulation of SLC37A1 in cancer cells , researchers should consider:

• Cancer type specificity: Focus on cancers with known EGFR pathway activation or phospholipid metabolism alterations

• Correlation with proliferation markers: Combine SLC37A1 immunodetection with Ki-67 or other proliferation markers

• Therapy response biomarkers: Investigate whether SLC37A1 expression changes correlate with response to EGFR-targeted therapies

• Metabolic profiling: Integrate SLC37A1 expression data with metabolomic profiling of glycerophospholipids and related metabolites

• Prognostic indicators: Evaluate potential correlations between SLC37A1 expression and clinical outcomes

Research has demonstrated that EGF transactivates the SLC37A1 promoter sequence and induces SLC37A1 mRNA and protein expression through the EGFR/MAPK/Fos pathway in ER-negative breast cancer cells, suggesting potential relevance as a biomarker or therapeutic target .