TMEM213 Antibody

What is TMEM213 and why is it relevant for research?

TMEM213 (Transmembrane protein 213) is an integral membrane protein encoded by the TMEM213 gene located on chromosome 7q34 in humans. It has a molecular weight of approximately 11,555 Da and exists in four isoforms produced by alternative splicing . Recent studies have identified TMEM213 as a potential prognostic biomarker in certain cancers, particularly lung adenocarcinoma, making it increasingly relevant for oncology research . The protein plays a role in cellular processes and signaling pathways, though its precise molecular function requires further elucidation. The growing interest in TMEM213 stems from its potential implications in disease progression and response to therapy, positioning it as an important research target for understanding membrane protein biology in normal and pathological conditions.

What are the key structural and functional characteristics of TMEM213?

TMEM213 is classified as an integral membrane protein with a cellular component designation as "integral to membrane" . The protein has multiple transmembrane domains, consistent with its role as a membrane-spanning protein. While the complete three-dimensional structure has not been fully resolved, sequence analysis indicates it contains specific functional domains.

The human TMEM213 protein (UniProt code: A2RRL7) shares significant sequence homology with mouse and rat orthologs (90% identity), suggesting evolutionary conservation of function across mammalian species . Four distinct isoforms exist due to alternative splicing, potentially conferring different functional properties. The protein's localization to the membrane suggests it may function in cellular signaling, transport, or membrane organization. Recent functional studies have begun exploring its role in cancer biology, particularly in relation to treatment response in lung adenocarcinoma, indicating a potential function in cellular pathways relevant to cancer progression and therapy response .

What types of TMEM213 antibodies are commercially available and what are their characteristics?

Several TMEM213 antibodies are commercially available with different specifications suitable for various research applications. The primary types include:

These antibodies are primarily rabbit polyclonal antibodies that recognize human TMEM213, though some show cross-reactivity with mouse and rat orthologs. The immunogens used for antibody production vary, with some targeting specific amino acid sequences and others using recombinant protein fragments. All are intended for research use only and not for diagnostic procedures .

What are the optimal experimental conditions for using TMEM213 antibodies in different applications?

The optimal experimental conditions for TMEM213 antibodies vary based on the specific application:

For Immunohistochemistry (IHC):

• Recommended dilution: 1:20-1:200 for PACO30566 and 1:50-1:200 for Novus Biologicals antibody

• Validated on: Paraffin-embedded human testis tissue

• Visualization: Typically using standard detection systems compatible with rabbit antibodies

• Protocol recommendation: Antigen retrieval may be necessary; specific buffer conditions should be optimized

For Immunofluorescence (IF):

• Recommended dilution: 1:50-1:200

• Validated on: HeLa cells

• Secondary antibody: Alexa Fluor 488-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) has been successfully used

• Buffer conditions: Standard IF blocking and washing buffers

For ELISA:

• Recommended dilution: 1:2000-1:10000

• Buffer conditions: Standard ELISA coating, blocking, and washing buffers

• Detection system: Compatible with rabbit IgG detection systems

Storage and handling recommendations for all TMEM213 antibodies include:

• Short-term storage at 4°C

• Long-term storage at -20°C with aliquoting to avoid freeze-thaw cycles

• Preservation in appropriate buffer (e.g., PBS pH 7.4 with 50% glycerol and preservatives)

Optimization of these conditions for specific experimental setups is recommended, as factors such as tissue type, fixation method, and detection system can influence results.

How can researchers validate the specificity of TMEM213 antibodies in their experimental systems?

Researchers can employ multiple validation approaches to ensure the specificity of TMEM213 antibodies:

1. Orthogonal Validation:

• Compare protein detection with mRNA expression data

• Perform correlation analysis between antibody staining and RNA-seq or qPCR data

• This approach evaluates consistency between protein and transcript levels

2. Independent Antibody Validation:

• Use multiple antibodies targeting different epitopes of TMEM213

• Compare staining patterns and localization

• Consistent results across different antibodies increase confidence in specificity

3. Genetic Approaches:

• Use TMEM213 knockout or knockdown models as negative controls

• Overexpression systems as positive controls

• CRISPR-Cas9 edited cell lines can provide definitive validation

4. Biochemical Validation:

• Perform western blotting to confirm molecular weight (expected ~11.5 kDa)

• Conduct immunoprecipitation followed by mass spectrometry

• Pre-adsorption with immunizing peptide should abolish specific signal

5. Cross-reactivity Assessment:

• Test antibody on tissues/cells known to express or not express TMEM213

• Examine reactivity with mouse/rat samples (90% sequence identity) to confirm cross-species specificity

6. Reliability Assessment Framework:

• Evaluate antibody according to established reliability criteria

• Aim for "Enhanced" validation status that requires multiple validation approaches

Comprehensive validation using multiple approaches provides the highest confidence in antibody specificity and experimental results.

What methodologies are recommended for studying TMEM213 expression in tissue samples?

For studying TMEM213 expression in tissue samples, several complementary methodologies are recommended:

Immunohistochemistry (IHC):

• Use validated dilution ranges (1:20-1:200) of TMEM213 antibodies

• Include appropriate positive controls (such as human testis tissue, which has been validated)

• Employ antigen retrieval methods optimized for membrane proteins

• Use automated staining platforms for consistency when possible

• Quantify expression using digital image analysis software for objectivity

Immunofluorescence (IF):

• Particularly useful for co-localization studies with other cellular markers

• Recommended dilution of 1:50-1:200

• Use confocal microscopy for precise subcellular localization

• Consider multi-color IF to investigate interaction with other proteins

RNA-based Methods:

• In situ hybridization to detect TMEM213 mRNA in tissue context

• RNAscope® technology for sensitive and specific mRNA detection

• qRT-PCR for quantitative assessment of TMEM213 transcript levels

• RNA-seq for comprehensive transcriptomic profiling

Tissue Microarrays (TMAs):

• Efficient for studying multiple tissue samples simultaneously

• Useful for determining expression patterns across different tissue types

• Can be combined with IHC or IF methods

Digital Spatial Profiling:

• Advanced technique for spatial characterization of TMEM213 expression

• Provides quantitative data on expression in the tissue microenvironment

• Allows multiplex analysis with other proteins of interest

A combined approach is often most informative, correlating protein expression (IHC/IF) with mRNA levels to provide validation and comprehensive expression profiling of TMEM213 in tissue samples.

What is the evidence for TMEM213 as a prognostic biomarker in cancer research?

Current evidence suggests TMEM213 may serve as a valuable prognostic biomarker, particularly in lung adenocarcinoma. Key findings include:

Prognostic Value in Lung Adenocarcinoma:

Association with Treatment Response:

Validation Approaches:

• Internal validation was conducted using bootstrapping methods (1,000 replications)

• External validation was performed using the online Kaplan-Meier plotter database

• These validation approaches strengthened the evidence for TMEM213's prognostic significance

Biological Pathway Associations:

While these findings are promising, further investigation is needed to fully establish TMEM213 as a clinically applicable biomarker. Additional studies with larger cohorts, diverse cancer types, and mechanistic investigations would strengthen the evidence base for TMEM213's utility as a prognostic and predictive biomarker in oncology.

How can researchers integrate TMEM213 expression data with other molecular markers for comprehensive pathway analysis?

Researchers can employ several sophisticated approaches to integrate TMEM213 expression data with other molecular markers for comprehensive pathway analysis:

1. Gene Set Enrichment Analysis (GSEA):

• Use GSEA software (such as GSEA v2.2.2) to analyze pathway enrichment based on TMEM213 expression levels

• Categorize samples into high and low TMEM213 expression groups

• Utilize curated gene sets from MSigDB, particularly KEGG pathway collections

• Identify significantly enriched pathways associated with TMEM213 expression patterns

2. Multi-omics Integration Approaches:

• Correlate TMEM213 protein expression (antibody-based detection) with:

  • mRNA expression (RNA-seq or microarray data)

  • Epigenetic profiles (methylation patterns, histone modifications)

  • Mutation data (whole-exome or targeted sequencing)

  • Protein interactome data (mass spectrometry-based proteomics)

• Employ computational tools specifically designed for multi-omics integration (e.g., iCluster, MOFA)

3. Network Analysis Methods:

• Construct protein-protein interaction networks including TMEM213

• Identify hub proteins and signaling modules connected to TMEM213

• Use tools like Cytoscape with plugins such as ReactomeFI or STRING

• Apply network medicine approaches to identify disease modules

4. Machine Learning Integration:

• Develop predictive models incorporating TMEM213 with other molecular markers

• Use techniques such as random forests, support vector machines, or deep learning

• Evaluate feature importance to determine TMEM213's relative contribution

• Perform cross-validation to ensure model robustness

5. Single-cell Analysis Integration:

• Examine TMEM213 expression at single-cell resolution

• Identify cell populations with distinctive TMEM213 expression patterns

• Integrate with other markers to define cell states and transitions

• Map TMEM213 to specific cellular programs or differentiation trajectories

6. Clinical-Molecular Correlations:

• Integrate TMEM213 expression with clinical parameters

• Perform multivariate analyses including:

  • Traditional clinicopathological parameters (age, gender, stage)

  • Other molecular markers

  • Treatment response data

• Use nomograms or other predictive tools to visualize integrated prognostic models

These integration approaches provide a comprehensive understanding of TMEM213's biological significance within the context of broader cellular pathways and disease mechanisms.

What experimental designs are recommended for investigating TMEM213's functional role in cellular processes?

To investigate TMEM213's functional role in cellular processes, researchers should consider the following experimental designs:

1. Gene Modulation Studies:

• Knockdown/Knockout Approaches:

  • siRNA or shRNA-mediated knockdown for temporary suppression

  • CRISPR-Cas9 gene editing for complete knockout

  • Inducible systems (e.g., Tet-On/Off) for temporal control of expression

• Overexpression Systems:

  • Transient transfection with tagged TMEM213 constructs

  • Stable cell lines with regulated TMEM213 expression

  • Domain-specific mutants to investigate structure-function relationships

2. Subcellular Localization and Trafficking:

• Advanced Microscopy:

  • Live-cell imaging with fluorescently tagged TMEM213

  • Super-resolution microscopy for detailed localization

  • FRAP (Fluorescence Recovery After Photobleaching) to assess mobility

• Biochemical Fractionation:

  • Membrane fractionation to confirm integral membrane localization

  • Density gradient centrifugation to identify specific membrane compartments

  • Surface biotinylation to quantify plasma membrane expression

3. Protein Interaction Studies:

• Co-immunoprecipitation:

  • Using validated TMEM213 antibodies to pull down interaction partners

  • Reverse co-IP with antibodies against suspected interaction partners

• Proximity Labeling:

  • BioID or APEX2 approaches to identify proximal proteins

  • Split-BioID for detecting specific interaction interfaces

• Yeast Two-Hybrid or Mammalian Two-Hybrid:

  • Screening for direct protein-protein interactions

4. Functional Assays Based on Cancer Biology Findings:

• Cell Proliferation Assays:

  • MTT, BrdU incorporation, or real-time cell analysis

  • Colony formation assays to assess long-term growth effects

• Migration and Invasion Assays:

  • Wound healing, transwell, and 3D invasion assays

• Apoptosis and Cell Death:

  • Annexin V/PI staining, caspase activation assays

• Drug Response Assays:

  • Dose-response studies with chemotherapeutic agents

  • Combination therapies to assess synergistic effects

5. Pathway Analysis:

• Reporter Assays:

  • Luciferase-based pathway reporters (e.g., MAPK, Wnt, NF-κB)

  • Phosphorylation-specific antibodies to assess signaling cascades

• Transcriptional Profiling:

  • RNA-seq following TMEM213 modulation

  • ChIP-seq to identify associated transcription factors

6. In Vivo Models:

• Xenograft Models:

  • Comparing growth of tumors with modified TMEM213 expression

  • Patient-derived xenografts to maintain tumor heterogeneity

• Genetically Engineered Mouse Models:

  • Conditional knockout/knockin of TMEM213

  • Tissue-specific expression systems

7. Clinical Correlation Studies:

• Tissue Microarrays:

  • Correlating TMEM213 expression with clinical outcomes

  • Multi-marker analysis for pathway activation

• Liquid Biopsy Approaches:

  • Examining TMEM213 in circulating tumor cells or exosomes

These experimental approaches provide a comprehensive framework for investigating TMEM213's functional roles, potentially revealing mechanisms behind its association with cancer prognosis and treatment response.

What are common technical challenges when working with TMEM213 antibodies and how can they be addressed?

Researchers working with TMEM213 antibodies may encounter several technical challenges. Here are common issues and recommended solutions:

1. Background Staining and Non-specific Binding:

• Problem: High background or non-specific staining in IHC or IF applications

• Solutions:

  • Optimize antibody dilution (test range from 1:20-1:200 for IHC and 1:50-1:200 for IF)

  • Increase blocking time and concentration (5% BSA or normal serum from secondary antibody host)

  • Include 0.1-0.3% Triton X-100 for membrane permeabilization

  • Use more stringent washing steps (increase number and duration)

  • Pre-adsorb antibody with non-specific proteins

2. Inconsistent Membrane Protein Detection:

• Problem: Variable or weak staining of TMEM213 as a membrane protein

• Solutions:

  • Optimize fixation methods (membrane proteins can be sensitive to overfixation)

  • Use gentler permeabilization methods

  • Try different antigen retrieval methods (citrate vs. EDTA-based)

  • Consider using fresh frozen tissues rather than paraffin-embedded samples

  • Test detergent concentration carefully to preserve membrane integrity

3. Antibody Specificity Concerns:

• Problem: Difficulty confirming antibody specificity

• Solutions:

  • Perform validation using multiple antibodies targeting different epitopes

  • Include proper controls (tissue known to express or lack TMEM213)

  • Conduct peptide competition assays with the immunizing peptide

  • Verify antibody reliability using orthogonal validation approaches

  • Consider using genetic knockdown/knockout as definitive controls

4. Storage and Stability Issues:

• Problem: Loss of antibody activity over time

• Solutions:

  • Store at 4°C short term only; aliquot and freeze at -20°C for long term

  • Avoid repeated freeze-thaw cycles by making small aliquots

  • Add carrier protein (BSA) for diluted antibody solutions

  • Check manufacturer's recommendations for specific storage buffers

5. Cross-Reactivity with Related Proteins:

• Problem: Antibody binds to proteins other than TMEM213

• Solutions:

  • Use antibodies validated for specificity across multiple proteins

  • Consider antibodies specifically tested on protein arrays

  • Perform western blot analysis to confirm detection at correct molecular weight (~11.5 kDa)

  • Validate across multiple species if conducting comparative studies

6. Reproducibility Issues:

• Problem: Inconsistent results between experiments

• Solutions:

  • Standardize protocols meticulously (timing, temperature, reagent concentrations)

  • Use automated staining platforms when possible

  • Prepare larger batches of working solutions

  • Document lot numbers and validate new antibody lots against previous results

  • Consider using positive control samples in each experimental run

Addressing these technical challenges requires systematic optimization and thorough controls, but implementing these solutions can significantly improve the reliability and reproducibility of TMEM213 antibody applications.

How should researchers approach contradictory data between TMEM213 protein expression and mRNA levels?

When researchers encounter contradictions between TMEM213 protein expression (detected by antibodies) and mRNA levels, a systematic analytical approach is necessary:

Understanding Potential Causes of Discrepancies:

• Post-transcriptional Regulation Mechanisms:

  • MicroRNA-mediated repression of translation

  • RNA-binding proteins affecting mRNA stability or translation efficiency

  • Alternative splicing generating different protein isoforms (TMEM213 has 4 known isoforms)

  • Differences in mRNA and protein half-lives

• Technical and Methodological Factors:

  • Antibody specificity or sensitivity limitations

  • Different detection thresholds between protein and mRNA methods

  • Sampling differences (spatial or temporal)

  • Fixation artifacts affecting epitope accessibility

Recommended Investigative Approaches:

• Validation of Both Measurement Methods:

  • Re-validate antibody specificity using orthogonal approaches

  • Confirm mRNA detection specificity with multiple primer sets or probes

  • Test multiple antibodies targeting different TMEM213 epitopes

  • Use positive and negative controls for both protein and mRNA detection

• Temporal Analysis:

  • Conduct time-course experiments to detect potential delays between transcription and translation

  • Measure mRNA and protein stability using actinomycin D and cycloheximide chase experiments

• Spatial Resolution Analysis:

  • Perform in situ hybridization alongside IHC on sequential tissue sections

  • Use single-cell approaches to detect cell-type-specific discrepancies

  • Consider subcellular localization affecting protein detection

• Regulatory Mechanism Investigation:

  • Perform polysome profiling to assess translation efficiency

  • Analyze miRNA expression profiles and test potential regulatory miRNAs

  • Investigate protein degradation rates and ubiquitination status

• Integrated Analysis Framework:

  • Calculate correlation coefficients between mRNA and protein levels across samples

  • Develop mathematical models accounting for synthesis and degradation rates

  • Use machine learning approaches to identify patterns in discrepancies

  • Apply statistical methods specifically designed for integration of multi-omics data

• Biological Validation:

  • Conduct functional experiments with gene modulation (overexpression/knockdown)

  • Monitor changes in both mRNA and protein levels following perturbation

  • Investigate response to relevant stimuli that might affect post-transcriptional regulation

It's important to note that genes not detected at the mRNA level but detected at the protein level (or vice versa) are present in all evidence categories, indicating this is a common biological phenomenon rather than simply experimental error . When reporting such discrepancies, researchers should clearly document the methodologies used, consider multiple explanations, and design follow-up experiments to investigate the underlying mechanisms rather than assuming error in either measurement.

What quality control parameters should be evaluated when selecting and using TMEM213 antibodies?

When selecting and using TMEM213 antibodies, researchers should evaluate several critical quality control parameters to ensure reliable and reproducible results:

1. Antibody Validation Documentation:

• Validation Method Classification:

  • Assess whether the antibody has Enhanced, Supported, or Approved validation status

  • Prioritize antibodies validated through multiple independent methods

• Validation Methodology:

  • Check for orthogonal validation (correlation with mRNA expression)

  • Look for validation using independent antibodies targeting different epitopes

  • Verify validation in specific applications (IHC, IF, ELISA, etc.)

• Specificity Testing:

  • Confirm testing on protein arrays containing target plus non-specific proteins

  • Check for cross-reactivity testing with related proteins

2. Technical Specifications:

• Immunogen Details:

  • Verify the exact sequence used for immunization (e.g., LLCVDKLMKL TPDEPKDLQA for some TMEM213 antibodies)

  • Assess whether the immunogen is conserved across species if cross-reactivity is needed

• Clonality and Source:

  • Note whether the antibody is monoclonal or polyclonal (most current TMEM213 antibodies are polyclonal)

  • Consider the host species (typically rabbit for available TMEM213 antibodies)

• Purification Method:

  • Check purification technique (e.g., Protein G purification, affinity purification)

  • Assess purity percentage (e.g., >95% for PACO30566)

3. Application-Specific Parameters:

• Recommended Dilutions:

  • Verify optimal dilutions for specific applications:

    • ELISA: 1:2000-1:10000

    • IHC: 1:20-1:200

    • IF: 1:50-1:200

• Validated Applications:

  • Confirm the antibody has been specifically tested for your application of interest

  • Check the specific conditions used in validation (tissue types, cell lines, etc.)

• Species Reactivity:

  • Verify reactivity with species of interest (human, mouse, rat)

  • Note percent homology with orthologs (e.g., 90% identity with mouse and rat)

4. Physical Properties and Handling:

• Storage Buffer Composition:

  • Note preservatives and stabilizers (e.g., 0.03% Proclin 300, 50% Glycerol, PBS pH 7.4)

  • Check for potentially interfering components for specific applications

• Storage Requirements:

  • Verify proper storage conditions (4°C short term, -20°C long term)

  • Note aliquoting recommendations to avoid freeze-thaw cycles

• Shelf Life:

  • Check expiration date and stability data

  • Consider lot-to-lot consistency for long-term studies

5. Performance Documentation:

• Positive Controls:

  • Identify validated positive control tissues (e.g., human testis for TMEM213)

  • Look for example images showing expected staining patterns

• Sensitivity Assessment:

  • Check detection limit information if available

  • Assess signal-to-noise ratio in example images

• Reproducibility Data:

  • Look for evidence of lot-to-lot consistency

  • Check for inter-laboratory validation if available

By systematically evaluating these quality control parameters, researchers can select TMEM213 antibodies with the highest likelihood of generating reliable, specific, and reproducible results in their experimental systems.

What are promising future research directions for TMEM213 in relation to disease mechanisms and therapeutic applications?

Based on current knowledge and preliminary findings, several promising research directions for TMEM213 warrant further investigation:

Cancer Biology and Therapeutic Applications:

• Expanded Biomarker Validation Studies:

  • Validate TMEM213's prognostic value across multiple cancer types beyond lung adenocarcinoma

  • Conduct larger prospective clinical studies to confirm predictive value for treatment response

  • Develop standardized diagnostic assays for clinical implementation

• Therapeutic Target Exploration:

  • Investigate whether modulating TMEM213 expression affects cancer cell sensitivity to chemotherapy

  • Explore the potential of TMEM213-targeted therapies (antibody-drug conjugates, small molecule modulators)

  • Study combination approaches with existing cancer treatments

Molecular Mechanism Investigations:

• Signaling Pathway Integration:

  • Map TMEM213's position in key cellular signaling networks

  • Identify TMEM213 interacting partners through proteomics approaches

  • Characterize post-translational modifications regulating TMEM213 function

• Structure-Function Analysis:

  • Determine the three-dimensional structure of TMEM213 protein

  • Map functional domains and critical amino acid residues

  • Investigate how the four identified isoforms differ in function

Physiological Role Elucidation:

• Tissue-Specific Functions:

  • Characterize TMEM213 expression patterns across normal tissues and developmental stages

  • Investigate tissue-specific regulatory mechanisms controlling TMEM213 expression

  • Develop tissue-specific knockout models to elucidate function in different organ systems

• Membrane Biology:

  • Explore TMEM213's role in membrane organization and dynamics

  • Investigate potential functions in cellular transport or ion channel regulation

  • Study subcellular localization and trafficking mechanisms

Emerging Technology Applications:

• Single-Cell Analysis:

  • Apply single-cell transcriptomics and proteomics to map TMEM213 expression at cellular resolution

  • Identify cell populations with distinctive TMEM213 expression patterns

  • Correlate with cell states and differentiation trajectories

• Advanced Imaging:

  • Utilize super-resolution microscopy to study TMEM213 distribution in membranes

  • Apply live-cell imaging to track dynamic changes in TMEM213 localization

  • Develop new TMEM213 antibody-based imaging probes

Translational Research:

• Liquid Biopsy Development:

  • Investigate TMEM213 detection in circulating tumor cells or exosomes

  • Develop minimally invasive monitoring approaches for treatment response

• Precision Medicine Applications:

  • Create predictive models incorporating TMEM213 status for treatment selection

  • Design clinical trials stratifying patients based on TMEM213 expression

  • Develop companion diagnostics for emerging TMEM213-targeted therapies

These research directions represent opportunities to transform our understanding of TMEM213 from a relatively understudied membrane protein to a well-characterized molecular entity with potential clinical applications in disease diagnosis, prognosis, and treatment.

How can emerging technologies enhance the study of TMEM213 expression and function?

Emerging technologies are revolutionizing protein research and offer significant advantages for studying TMEM213 expression and function:

Advanced Proteomics Technologies:

• Mass Spectrometry Innovations:

  • Targeted proteomics (SRM/MRM) for precise quantification of TMEM213 without antibody dependency

  • Top-down proteomics for characterizing intact TMEM213 protein and its isoforms

  • Cross-linking mass spectrometry (XL-MS) to map TMEM213 interaction networks in native membrane environments

• Proximity Labeling Technologies:

  • BioID, APEX2, or TurboID systems fused to TMEM213 to identify proximal proteins

  • Spatially-resolved proximity labeling to map interaction networks in specific cellular compartments

  • Split-proximity labeling for detecting specific protein-protein interactions

Advanced Genomic and Transcriptomic Technologies:

• CRISPR-Based Technologies:

  • CRISPR interference/activation for precise modulation of TMEM213 expression

  • CRISPR screens to identify genetic interactions with TMEM213

  • Base editing and prime editing for introducing specific mutations

• Spatial Transcriptomics:

  • Visium or Slide-seq for spatial mapping of TMEM213 mRNA in tissue context

  • Integration with protein detection methods for multi-modal spatial analysis

  • Single-cell spatial transcriptomics for cellular heterogeneity assessment

Advanced Imaging Technologies:

• Super-Resolution Microscopy:

  • STORM, PALM, or STED microscopy for nanoscale visualization of TMEM213 in membranes

  • Multi-color super-resolution for co-localization with interaction partners

  • Live-cell super-resolution for dynamic studies

• Correlative Light and Electron Microscopy (CLEM):

  • Precise localization of TMEM213 in ultrastructural context

  • Immuno-EM for high-resolution localization studies

• Expansion Microscopy:

  • Physical expansion of specimens for enhanced resolution with standard microscopes

  • Compatible with multiple rounds of staining for multiplexed analysis

Microfluidics and Organ-on-Chip Technologies:

• Cell Culture Advances:

  • Microfluidic organ-on-chip systems for studying TMEM213 in physiologically relevant conditions

  • 3D organoid cultures for examining TMEM213 function in tissue-like environments

  • Patient-derived organoids to study TMEM213 in disease contexts

• Single-Cell Analysis:

  • Droplet-based single-cell proteomics for TMEM213 quantification at cellular resolution

  • Single-cell western blotting for protein heterogeneity assessment

  • Microfluidic antibody capture for single-cell surface protein analysis

Computational and AI-Based Approaches:

• Protein Structure Prediction:

  • AlphaFold2 and RoseTTAFold for TMEM213 structure prediction

  • Molecular dynamics simulations of TMEM213 in membrane environments

  • Virtual screening for identifying molecules interacting with TMEM213

• Multi-Omics Integration:

  • Machine learning models integrating TMEM213 data across multiple platforms

  • Network analysis tools for placing TMEM213 in biological pathways

  • Digital pathology with AI analysis of TMEM213 staining patterns

Antibody Engineering and Alternatives:

• Next-Generation Antibodies:

  • Nanobodies or single-domain antibodies for improved access to membrane protein epitopes

  • Recombinant antibody fragments with site-specific conjugation for improved consistency

• Aptamer Technology:

  • DNA/RNA aptamers as alternatives to antibodies for TMEM213 detection

  • Cell-SELEX for generating aptamers against native TMEM213 in cellular context

These emerging technologies can overcome limitations of traditional approaches and provide unprecedented insights into TMEM213 biology, potentially accelerating both fundamental understanding and clinical applications.