SLC2A2 Recombinant Monoclonal Antibody

What is SLC2A2/GLUT2 and what is its biological significance?

SLC2A2 (Solute Carrier Family 2 Member 2), also known as GLUT2 (Glucose Transporter Type 2), is a 524-amino acid membrane-associated protein that functions as the principal transporter for glucose transfer between liver and blood. It enables protein-facilitated glucose movement across cell membranes and plays a crucial role in maintaining whole-body glucose homeostasis . The protein belongs to the Major Facilitator Superfamily, specifically the Sugar Transporter (TC 2.A.1.1) family, Glucose Transporter subfamily . SLC2A2 is essential for glucose uptake and utilization in tissues such as the liver, pancreas, and intestine, with dysregulation being implicated in metabolic disorders including diabetes and obesity .

What is the molecular structure and cellular localization of SLC2A2?

SLC2A2 is a multi-pass membrane protein with a calculated molecular weight of 57kDa. The protein contains glycosylation sites and has been mapped with several functional domains . A partial amino acid sequence of human SLC2A2 includes: "MTED KVTG TLVF TVIT AVLG SFQF GYDI GVIN APQQ VIIS HYRH VLGV PLDD RKAI NNYV INST DELP TISY SMNP KPTP WAEE ETVA AAQL ITML WSLS" . The protein is predominantly localized to the plasma membrane, where it facilitates bidirectional glucose transport across the cell boundary .

How are SLC2A2 Recombinant Monoclonal Antibodies produced?

SLC2A2 recombinant monoclonal antibodies are synthetically produced through a systematic in vitro approach. The process begins with the extraction of SLC2A2 antibody genes from B cells isolated from immunoreactive rabbits. These genes undergo amplification and are cloned into suitable phage vectors, which are subsequently introduced into mammalian cell lines to facilitate the production of functional antibodies in significant quantities. The antibodies are then purified using affinity chromatography techniques . This recombinant production method ensures higher batch-to-batch consistency compared to traditional hybridoma-derived monoclonal antibodies.

What applications are SLC2A2 Recombinant Monoclonal Antibodies validated for?

SLC2A2 Recombinant Monoclonal Antibodies have been validated for multiple research applications, as outlined in the following table:

These applications enable researchers to detect human SLC2A2 protein in various experimental contexts .

What positive controls are recommended for validating SLC2A2 antibody specificity?

When validating the specificity of SLC2A2 antibodies, researchers should consider using the following positive control samples that are known to express SLC2A2: HT-29 cells, K-562 cells, rat liver tissue, and rat kidney tissue . Additionally, recombinant SLC2A2 proteins with greater than 85% purity (as determined by SDS-PAGE) can serve as reliable positive controls for antibody validation experiments . Incorporating these controls helps establish antibody specificity and provides a benchmark for expected signal intensity in experimental samples.

How should sample preparation be optimized for SLC2A2 detection in membrane fractions?

For optimal detection of SLC2A2 in membrane fractions, researchers should implement a multi-step protocol that preserves membrane integrity while maximizing protein extraction efficiency. Begin with gentle cell lysis using a non-ionic detergent buffer (e.g., containing 1% Triton X-100 or 0.5% NP-40) supplemented with protease inhibitors. Differential centrifugation should follow, with an initial low-speed centrifugation (500-1000×g) to remove nuclei and cell debris, followed by high-speed ultracentrifugation (100,000×g for 1 hour) to isolate membrane fractions.

For Western blotting applications, samples should not be boiled as this may cause aggregation of membrane proteins; instead, incubate at 37°C for 30 minutes in sample buffer. When processing tissue samples, cryosectioning followed by methanol fixation has shown superior results for preserving SLC2A2 epitopes compared to formalin fixation . These methodological considerations are essential as improper sample preparation can significantly impact antibody binding and detection sensitivity.

How can SLC2A2 Recombinant Monoclonal Antibodies be used to study the dual function of GLUT2 as both transporter and receptor?

Research has revealed that SLC2A2/GLUT2 functions not only as a glucose transporter but also as a receptor involved in signaling pathways. To investigate this dual functionality, researchers can employ SLC2A2 Recombinant Monoclonal Antibodies in conjunction with structure-function analyses of wild-type and mutant GLUT2 proteins. One effective approach involves characterizing a panel of mutations along the protein and assessing their differential impact on transport versus receptor activity .

The methodology should include:

• Expressing wild-type and mutant SLC2A2 constructs (with HA tags for detection) in appropriate cell models

• Using SLC2A2 antibodies to confirm membrane expression through immunofluorescence and flow cytometry

• Assessing glucose transport kinetics in systems like Xenopus oocytes

• Evaluating receptor-mediated functions such as glucose-induced insulin secretion

• Analyzing downstream signaling activation through phosphorylation studies

This research strategy has been successfully implemented to identify specific amino acids differentially involved in the two hGLUT2 functions, as demonstrated in studies of naturally occurring SLC2A2 variants and engineered mutations based on sequence alignments .

What insights can SLC2A2 antibody-based studies provide about Fanconi-Bickel syndrome?

Fanconi-Bickel syndrome (FBS) is a rare genetic disorder caused by inactivating mutations in the SLC2A2 gene. SLC2A2 Recombinant Monoclonal Antibodies serve as valuable tools for investigating the molecular pathology of this condition. By employing these antibodies in combination with site-directed mutagenesis and functional assays, researchers can characterize how FBS-associated mutations affect protein expression, localization, and function.

Studies using SLC2A2 antibodies have revealed that four proposed inactivating mutations associated with FBS significantly impact glucose transport and insulin secretion . The experimental approach involves:

• Generating constructs with FBS-associated mutations through site-directed mutagenesis

• Transfecting these constructs into relevant cell models

• Using SLC2A2 antibodies to assess membrane expression in hepatic and pancreatic β cells

• Performing transport kinetics assays in Xenopus oocytes

• Evaluating glucose-induced insulin secretion

• Analyzing effects on pancreatic β cell development

These comprehensive analyses have refined the structure-function map of hGLUT2, highlighting the importance of its sugar receptor activity and identifying it as a potential target for stimulating pancreatic β cell differentiation and insulin secretion .

How can SLC2A2 Recombinant Monoclonal Antibodies be employed in studying metabolic disorders?

SLC2A2 Recombinant Monoclonal Antibodies provide valuable tools for investigating the role of GLUT2 in various metabolic disorders. Several research approaches have demonstrated their utility:

• Diabetes research: Antibodies can detect changes in GLUT2 expression in pancreatic β-cells following treatment with potential therapeutic compounds. For example, studies have shown that Rosiglitazone (RGZ) stimulates insulin release and synthesis through upregulation of GLUT-2, GCK, and BETA2/NeuroD gene expression .

• Obesity studies: By immunohistochemical analysis of liver and pancreatic tissues from obese models, researchers can assess alterations in GLUT2 distribution and expression levels.

• Natural compound effects: SLC2A2 antibodies have been used to demonstrate that compounds like p-Coumaric acid (p-CA) modulate glucose and lipid metabolism via GLUT2 activation in the pancreas, potentially offering beneficial effects for treating metabolic disorders .

• Glucose sensing mechanisms: Immunofluorescence studies using these antibodies have helped elucidate how functional β-cells respond to increased glucose levels by triggering insulin secretion, a process dependent on proper GLUT2 function .

By combining SLC2A2 antibody detection with functional assessments, researchers can gain comprehensive insights into the mechanistic basis of metabolic disorders and identify potential therapeutic targets.

What factors might affect the binding efficiency of SLC2A2 Recombinant Monoclonal Antibody?

Multiple factors can influence the binding efficiency of SLC2A2 Recombinant Monoclonal Antibodies, potentially impacting experimental outcomes. Understanding these factors is crucial for optimizing detection protocols:

• Epitope accessibility: The multi-pass membrane nature of SLC2A2 means that certain epitopes may be obscured by the membrane or by protein conformation. Different fixation and permeabilization methods can significantly alter epitope accessibility .

• Glycosylation status: SLC2A2 contains glycosylation sites that may interfere with antibody binding if they overlap with the target epitope. Sample preparation methods that affect glycosylation (e.g., enzymatic deglycosylation) may alter binding efficiency .

• Protein denaturation: For antibodies targeting conformational epitopes, the degree of protein denaturation during sample preparation is critical. Western blotting typically involves denatured proteins, while immunohistochemistry and immunofluorescence may preserve native conformations .

• Sample buffer composition: The presence of detergents, salts, or reducing agents in sample buffers can affect antibody-antigen interactions. Optimize buffer conditions based on the specific application and antibody characteristics.

• Cross-reactivity: While recombinant monoclonal antibodies offer high specificity, potential cross-reactivity with other glucose transporter family members should be considered, especially when working with complex samples .

By systematically evaluating these factors, researchers can troubleshoot binding issues and optimize experimental conditions for maximum detection sensitivity and specificity.

How should researchers interpret variations in SLC2A2 expression across different disease states?

Interpreting variations in SLC2A2 expression across disease states requires careful consideration of multiple factors. When analyzing data generated using SLC2A2 Recombinant Monoclonal Antibodies, researchers should:

By integrating these analytical approaches, researchers can derive meaningful insights from SLC2A2 expression patterns across different disease states and potentially identify new diagnostic or therapeutic targets.

What role does SLC2A2 play in cancer metabolism and how can recombinant antibodies facilitate this research?

• Tumor tissue profiling: Immunohistochemical analysis of tumor biopsies can reveal alterations in SLC2A2 expression patterns that correlate with tumor grade, stage, or metabolic phenotype.

• Metabolic flux analysis: By combining SLC2A2 antibody-based detection with glucose uptake assays, researchers can assess the relationship between transporter expression and functional glucose metabolism in cancer cells.

• Response to therapy: Monitoring changes in SLC2A2 expression following treatment with metabolic-targeting agents can provide insights into therapeutic mechanisms and potential resistance pathways.

• Cancer cell subtypes: Flow cytometry using SLC2A2 antibodies can help identify and isolate cancer cell populations with distinct metabolic profiles, enabling more detailed characterization of tumor heterogeneity.

This research direction holds promise for identifying new therapeutic targets and prognostic markers in cancers that exhibit altered glucose metabolism.

How can SLC2A2 Recombinant Monoclonal Antibodies contribute to understanding pancreatic β-cell differentiation and function?

SLC2A2 plays a critical role in pancreatic β-cell differentiation and glucose-stimulated insulin secretion. SLC2A2 Recombinant Monoclonal Antibodies offer powerful tools for investigating these processes through several methodological approaches:

• Developmental studies: Immunostaining of pancreatic tissue during different developmental stages can reveal the temporal pattern of SLC2A2 expression and its correlation with β-cell maturation.

• Stem cell differentiation: Monitoring SLC2A2 expression during directed differentiation of stem cells into insulin-producing cells provides a marker for β-cell identity and maturity.

• Functional β-cell assessment: Research has shown that functional β-cells respond to increased glucose levels by increasing insulin secretion, a process dependent on proper GLUT2 function. Antibody-based detection can help assess this functionality .

• Effects of mutations: Studies using antibodies to detect wild-type and mutant forms of SLC2A2 have demonstrated that mutations affecting glucose transport also impact pancreatic β-cell differentiation and insulin secretion .

• Therapeutic compound screening: Compounds like Rosiglitazone (RGZ) have been shown to stimulate insulin release and synthesis through upregulation of GLUT-2. Antibody-based detection can help screen additional compounds with similar effects .

These research applications highlight the potential of SLC2A2 as a target for stimulating pancreatic β-cell differentiation and insulin secretion, offering new therapeutic possibilities for diabetes management .

What are the emerging applications of SLC2A2 Recombinant Monoclonal Antibodies in precision medicine?

The development of highly specific SLC2A2 Recombinant Monoclonal Antibodies opens new avenues for precision medicine approaches targeting glucose metabolism disorders. Future applications may include:

• Personalized diabetes management: Analyzing SLC2A2 expression and function in patient samples could help stratify diabetic patients and guide personalized treatment strategies.

• Biomarker development: Given its association with clinical outcomes in conditions like hepatocellular carcinoma, SLC2A2 could serve as a biomarker for disease progression and treatment response .

• Targeted drug delivery: Antibody-drug conjugates targeting SLC2A2 could provide tissue-specific delivery of therapeutic agents to cells with high GLUT2 expression.

• Functional diagnostics: Assays incorporating SLC2A2 antibodies could help identify functional defects in glucose sensing and transport that contribute to metabolic disorders.

As our understanding of the dual transporter-receptor function of SLC2A2 continues to evolve, so too will the applications of these recombinant antibodies in both research and clinical settings .

What methodological advances are anticipated in SLC2A2 antibody development and application?

The field of SLC2A2 antibody development and application is likely to see several methodological advances in the coming years:

• Single-domain antibodies: Development of smaller antibody formats that can access epitopes within the transmembrane regions of SLC2A2, potentially offering new insights into structure-function relationships.

• Multiplex imaging: Combination of SLC2A2 antibodies with other metabolic markers in multiplex imaging approaches to provide comprehensive metabolic profiling at the single-cell level.

• Live-cell imaging: Development of non-disruptive antibody-based probes for tracking SLC2A2 dynamics in living cells, offering insights into transporter trafficking and regulation.

• Cryo-EM applications: Use of antibodies as tools for structure determination of SLC2A2 in different conformational states via cryo-electron microscopy.

• High-throughput screening: Implementation of antibody-based assays in high-throughput screening platforms to identify compounds that modulate SLC2A2 expression or function.