MS4A6A Antibody

What is MS4A6A and what is its biological significance?

MS4A6A (membrane spanning 4-domains A6A) is a protein belonging to the MS4A gene family that includes CD20 (MS4A1), FcRbeta (MS4A2), and Htm4 (MS4A3), along with at least 13 other genes encoding membrane proteins . Most MS4A family members display a characteristic tetraspanning topology and similar intron/exon splice boundaries . Functionally, MS4A6A is believed to participate in signal transduction as a component of a multimeric receptor complex . Recent research has identified MS4A6A as being associated with aging and the onset of neurodegenerative diseases, particularly Alzheimer's disease . Additionally, it has been implicated in cancer biology, specifically showing upregulation in glioma tissues, which correlates with unfavorable clinical outcomes and poor responses to adjuvant chemotherapy .

How is MS4A6A expression regulated in normal and pathological conditions?

In normal tissues, MS4A6A displays unique expression patterns among hematopoietic cells and non-lymphoid tissues, similar to other MS4A family members . Recent single-cell RNA sequencing (scRNA-seq) analyses have revealed cell-type specific expression patterns, particularly in immune cell populations . In pathological conditions, MS4A6A expression appears to be dysregulated through various mechanisms. For instance, in glioma, analysis of methylation patterns suggests potential epigenetic regulation involving DNA methyltransferases (DNMTs) such as DNMT1, DNMT3A, and DNMT3B . This epigenetic dysregulation contributes to abnormal MS4A6A expression levels that correlate with disease progression. In neurodegenerative contexts, alterations in MS4A6A expression have been linked to aging-related processes and may contribute to disease pathogenesis .

What criteria should be considered when selecting an MS4A6A antibody for research?

When selecting an MS4A6A antibody, researchers should consider several important criteria to ensure experimental success:

• Antibody Type: Choose between polyclonal and monoclonal antibodies based on your experimental needs. Polyclonal antibodies typically offer broader epitope recognition but may have batch-to-batch variability, while monoclonal antibodies provide consistent specificity but may be more sensitive to epitope masking .

• Species Reactivity: Verify that the antibody reacts with your species of interest. Currently available MS4A6A antibodies typically demonstrate reactivity with human and rat samples, but reactivity with other species may vary .

• Intended Applications: Confirm the antibody has been validated for your specific application. MS4A6A antibodies are commonly validated for Western blot (WB), ELISA, immunocytochemistry/immunofluorescence (ICC/IF), and immunohistochemistry on paraffin-embedded tissues (IHC-p) .

• Isoform Detection: Be aware that MS4A6A exists in at least three isoforms, and most antibodies detect only the two longer isoforms . If your research focuses on a specific isoform, select an antibody with appropriate epitope recognition.

• Conjugation: Determine whether you need an unconjugated antibody or one with a specific label/conjugate (e.g., fluorescent tags such as mFluor Violet 610 SE or DyLight 405) based on your detection methods .

How can researchers validate the specificity of an MS4A6A antibody?

Validating antibody specificity is critical for reliable experimental results. For MS4A6A antibodies, consider implementing these validation approaches:

• Western Blot Analysis: Confirm the antibody detects bands of the expected molecular weight (approximately 27 kDa for the predicted size, though observed size may be around 68 kDa) . Compare band patterns in samples with known differential expression of MS4A6A.

• Peptide Competition Assays: Pre-incubate the antibody with the immunizing peptide before application to samples. Specific binding should be blocked, resulting in diminished or absent signal .

• Positive and Negative Controls: Include tissue or cell samples known to express (e.g., certain immune cells) or not express MS4A6A.

• Cross-Reactivity Testing: Verify the antibody does not cross-react with other MS4A family members. Most commercial MS4A6A antibodies are designed to avoid cross-reactivity with related proteins .

• Genetic Validation: When possible, use samples from knockdown/knockout models or cells transfected with MS4A6A expression constructs to confirm antibody specificity.

• Orthogonal Techniques: Compare protein detection results with mRNA expression data from RT-PCR or RNA-seq to ensure concordance between transcript and protein levels.

What controls are essential when using MS4A6A antibodies in research applications?

Implementing appropriate controls is fundamental for obtaining reliable results with MS4A6A antibodies:

• Positive Control: Include samples known to express MS4A6A, such as certain immune cell populations or cell lines with confirmed MS4A6A expression .

• Negative Control: Use samples where MS4A6A expression is absent or samples from MS4A6A knockout models when available.

• Isotype Control: Include an irrelevant antibody of the same isotype, host species, and concentration as the MS4A6A antibody to identify potential non-specific binding.

• Secondary Antibody Only Control: Omit the primary MS4A6A antibody but include the secondary antibody to identify any background or non-specific binding from the secondary antibody.

• Blocking Peptide Control: When available, include a sample where the MS4A6A antibody has been pre-incubated with its immunogenic peptide to demonstrate binding specificity .

• Concentration Gradient Testing: For quantitative applications, establish a standard curve using different antibody concentrations to identify the optimal working concentration and ensure you're operating within the linear detection range .

What are the optimal conditions for using MS4A6A antibodies in Western blot applications?

For successful Western blot detection of MS4A6A, researchers should consider the following methodological guidelines:

• Sample Preparation: Extract proteins using buffers containing appropriate detergents for membrane proteins. RIPA buffer with protease inhibitors is often suitable for MS4A6A extraction.

• Protein Loading: Load 15-20 μg of total protein extract per lane, as demonstrated in published protocols .

• Antibody Concentration: Titrate the antibody to determine optimal concentration. Published protocols suggest using 1-2 μg/mL as a starting point for MS4A6A detection .

• Expected Molecular Weight: While the predicted molecular weight is approximately 27 kDa, be aware that MS4A6A may appear at around 68 kDa in some systems due to post-translational modifications or other factors .

• Detection Method: The ECL (enhanced chemiluminescence) technique has been successfully used for detecting MS4A6A in Western blots .

• Blocking Solution: Use 5% non-fat dry milk or BSA in TBST for blocking non-specific binding sites.

• Incubation Conditions: For primary antibody incubation, overnight at 4°C often yields optimal results, while secondary antibody incubation can be performed at room temperature for 1-2 hours.

• Membrane Type: PVDF membranes are generally recommended for optimal protein transfer and antibody binding.

How should researchers optimize immunohistochemistry protocols for MS4A6A detection in tissue sections?

For effective immunohistochemical detection of MS4A6A in tissue sections, consider these methodological recommendations:

• Tissue Preparation: Formalin-fixed, paraffin-embedded (FFPE) tissue sections are commonly used for MS4A6A detection. Proper fixation is critical for preserving antigen structure while maintaining tissue morphology.

• Antigen Retrieval: Perform heat-induced epitope retrieval using 10 mM sodium citrate buffer (pH 6.0) as documented in published protocols . This step is crucial for unmasking epitopes that may be cross-linked during fixation.

• Antibody Dilution: Start with the manufacturer's recommended dilution ratio (typically 1:200 for MS4A6A antibodies) and optimize as needed .

• Incubation Conditions: Incubate with primary antibody overnight at 4°C to maximize specific binding while minimizing background signal .

• Detection System: For chromogenic detection, use an appropriate HRP-labeled secondary antibody (typically at 1:200 dilution) followed by a suitable substrate such as DAB . For fluorescence detection, use appropriately labeled secondary antibodies and include DAPI for nuclear counterstaining.

• Multi-label Approaches: For co-localization studies, MS4A6A antibodies can be effectively combined with markers such as CD68 (macrophage marker) and CD163 (M2 macrophage marker) to investigate cell-specific expression patterns .

• Controls: Include positive controls (tissues known to express MS4A6A), negative controls (tissues not expressing MS4A6A), and technical controls (primary antibody omission) in each experiment.

• Image Acquisition: Use appropriate microscopy techniques (brightfield for chromogenic or fluorescence for immunofluorescence) with consistent exposure settings across samples to enable quantitative comparisons.

What are the key considerations for immunocytochemistry/immunofluorescence using MS4A6A antibodies?

When performing immunocytochemistry or immunofluorescence with MS4A6A antibodies, researchers should address these methodological aspects:

• Cell Fixation: Use 4% paraformaldehyde for 15-20 minutes at room temperature to preserve cell morphology while maintaining antigen accessibility.

• Permeabilization: Since MS4A6A is a membrane protein with intracellular domains, use a mild detergent (0.1-0.3% Triton X-100 or 0.1% saponin) for permeabilization to allow antibody access to all epitopes.

• Blocking: Block non-specific binding with 5-10% normal serum (from the species in which the secondary antibody was raised) in PBS containing 0.1-0.3% Triton X-100 for 1 hour at room temperature.

• Antibody Dilution and Incubation: Dilute MS4A6A antibodies according to manufacturer recommendations (typically 1:100 to 1:500) and incubate overnight at 4°C .

• Washing Steps: Perform thorough washing (3-5 times, 5 minutes each) with PBS between each step to reduce background and non-specific staining.

• Signal Detection: For fluorescent detection, select appropriate secondary antibodies conjugated to fluorophores compatible with your microscopy system. Alternatively, use directly conjugated MS4A6A antibodies (e.g., mFluor Violet 610 SE or DyLight 405) for direct detection .

• Counterstaining: Include DAPI (5 minutes in the dark) for nuclear counterstaining to facilitate cell identification and provide context for MS4A6A localization .

• Mounting: Use anti-fade mounting media to preserve fluorescence signal during imaging and storage.

• Image Acquisition: Utilize a fluorescence microscope with appropriate filter sets for the selected fluorophores, ensuring consistent exposure settings across all experimental conditions .

How can MS4A6A antibodies be utilized in single-cell analysis techniques?

MS4A6A antibodies can be valuable tools in various single-cell analysis approaches:

• Single-Cell Immunophenotyping: MS4A6A antibodies can be incorporated into multiparameter flow cytometry or mass cytometry (CyTOF) panels to identify and characterize specific cell populations expressing MS4A6A, particularly in immune cell subsets.

• Integration with Single-Cell RNA-Seq Data: Protein-level detection using MS4A6A antibodies can complement scRNA-seq data to validate gene expression findings at the protein level. Recent studies have used scRNA-seq to analyze MS4A6A expression in glioma samples, identifying cell-type specific expression patterns that could be further validated with antibody-based approaches .

• Imaging Mass Cytometry (IMC): MS4A6A antibodies can be metal-tagged for use in IMC to visualize spatial distribution of MS4A6A-expressing cells in tissue contexts while simultaneously detecting multiple other markers.

• CODEX Multiplexed Imaging: Conjugated MS4A6A antibodies can be incorporated into CODEX (CO-Detection by indEXing) panels for highly multiplexed imaging of tissues with single-cell resolution.

• Spatial Transcriptomics Integration: Combine immunofluorescence using MS4A6A antibodies with spatial transcriptomics techniques to correlate protein expression with transcriptional profiles in a spatially resolved manner.

• Live-Cell Imaging: Use non-blocking fluorescently conjugated MS4A6A antibodies to track dynamics of MS4A6A-expressing cells in real-time imaging experiments.

• Analysis Considerations: When analyzing single-cell data, implement appropriate computational tools for clustering, dimensionality reduction (e.g., UMAP, t-SNE), and trajectory analysis to identify patterns in MS4A6A expression across cell populations and states .

What approaches can address inconsistent or contradictory results when using MS4A6A antibodies?

When facing inconsistent or contradictory results with MS4A6A antibodies, consider implementing these troubleshooting approaches:

• Antibody Validation Revisit: Re-validate antibody specificity using orthogonal methods. Compare results from multiple antibody clones targeting different MS4A6A epitopes to increase confidence in observations.

• Isoform-Specific Analysis: Consider that MS4A6A exists in multiple isoforms, and different antibodies may preferentially detect specific isoforms . Verify which isoforms your antibody detects and design experiments to disambiguate isoform-specific effects.

• Post-Translational Modifications: Investigate whether post-translational modifications affect epitope accessibility or antibody binding. The discrepancy between predicted (27 kDa) and observed (68 kDa) molecular weights suggests potential glycosylation or other modifications that might influence detection .

• Protocol Optimization: Systematically optimize experimental conditions including fixation methods, antigen retrieval techniques, antibody concentration, and incubation conditions for each application.

• Technical Replicates: Perform multiple technical replicates to distinguish between biological variability and technical artifacts.

• Quantitative Analysis: Implement objective quantification methods rather than relying on subjective visual assessment. For imaging data, use appropriate software for unbiased quantification of signal intensity and localization patterns.

• Integrated Multi-Omics Approach: Combine antibody-based detection with other methodologies such as RNA-seq, proteomics, or genetic manipulation to build a more comprehensive understanding of MS4A6A biology.

• Tissue and Sample Context: Consider that MS4A6A expression and localization may vary based on tissue type, disease state, or experimental conditions. Context-specific regulation may explain apparently contradictory results across different systems.

How can MS4A6A antibodies be employed in studying neurodegenerative diseases and cancer?

MS4A6A antibodies offer valuable tools for investigating the protein's role in both neurodegenerative diseases and cancer:

• Neurodegenerative Disease Applications:

  • Biomarker Development: Validate MS4A6A as a potential biomarker for neurodegenerative diseases, particularly Alzheimer's disease, using antibody-based detection in patient samples .

  • Cell-Type Specific Expression: Use MS4A6A antibodies in combination with neuronal, microglial, and astrocytic markers to characterize cell-type specific expression patterns in healthy and diseased brain tissue.

  • Aging-Related Changes: Investigate age-dependent alterations in MS4A6A expression and localization using antibody-based detection in time-course studies.

  • Therapeutic Target Validation: Employ MS4A6A antibodies to validate the protein as a potential therapeutic target by assessing its expression, localization, and interaction partners in disease models.

• Cancer Research Applications:

  • Prognostic Marker Validation: Utilize MS4A6A antibodies in tissue microarrays or patient samples to validate MS4A6A as a prognostic biomarker, particularly in glioma where it has been associated with unfavorable clinical outcomes .

  • Tumor Microenvironment Characterization: Combine MS4A6A antibodies with macrophage markers (e.g., CD68, CD163) in multiplexed immunofluorescence to investigate the role of MS4A6A-expressing cells in the tumor microenvironment .

  • Therapy Response Prediction: Assess MS4A6A expression pre- and post-treatment to determine its utility as a predictive biomarker for therapy response, particularly for adjuvant chemotherapy in glioma patients .

  • Mechanistic Studies: Use MS4A6A antibodies in co-immunoprecipitation and proximity ligation assays to identify interaction partners and signaling pathways influenced by MS4A6A in cancer cells.

  • Immune Infiltrate Analysis: Apply MS4A6A antibodies in flow cytometry or multiplexed imaging to characterize immune infiltrates in tumors, potentially identifying targetable immune cell populations.

• Methodological Approaches:

  • Patient-Derived Models: Validate findings from patient samples in patient-derived xenografts or organoids using MS4A6A antibodies to track expression in more controlled experimental systems.

  • Longitudinal Studies: Implement MS4A6A antibody-based detection in longitudinal samples to track changes in expression during disease progression or treatment response.

  • Multiparameter Analysis: Combine MS4A6A detection with assessment of related signaling molecules to build comprehensive pathway maps in disease contexts.

How might MS4A6A antibodies be integrated with emerging single-cell and spatial biology technologies?

The integration of MS4A6A antibodies with cutting-edge technologies offers exciting opportunities for deeper biological insights:

• Spatial Proteomics: Incorporate MS4A6A antibodies into multiplexed imaging technologies such as CODEX, MIBI-TOF (Multiplexed Ion Beam Imaging by Time of Flight), or Vectra Polaris to analyze the spatial distribution of MS4A6A-expressing cells relative to other cell types and microenvironmental features.

• Antibody-Based Single-Cell Proteomics: Utilize MS4A6A antibodies in emerging single-cell proteomic techniques such as CITE-seq (Cellular Indexing of Transcriptomes and Epitopes by Sequencing) or REAP-seq (RNA Expression and Protein Sequencing) to simultaneously measure MS4A6A protein expression alongside transcriptomic profiles at single-cell resolution.

• Live-Cell Dynamics: Apply non-perturbing MS4A6A antibody fragments or nanobodies in live-cell imaging approaches to track dynamic changes in MS4A6A localization and interactions in real-time.

• Proximity Labeling: Combine MS4A6A antibodies with proximity labeling techniques such as BioID or APEX to identify the protein's interactome in different cellular contexts.

• Super-Resolution Microscopy: Employ fluorescently-labeled MS4A6A antibodies in STORM, PALM, or STED microscopy to visualize nanoscale organization and clustering of MS4A6A in membrane microdomains.

• Intravital Imaging: Utilize fluorescently-labeled MS4A6A antibodies for intravital microscopy to observe MS4A6A-expressing cells in living animals, particularly in disease models.

• Spatial Transcriptomics Integration: Correlate MS4A6A protein expression detected by antibodies with spatial transcriptomic data to build integrated maps of gene and protein expression in tissue contexts.

• Organ-on-Chip Models: Apply MS4A6A antibodies in organ-on-chip systems to investigate the protein's role in complex multicellular interactions under physiologically relevant conditions.

What methodological advances could improve MS4A6A detection in challenging samples?

Several methodological innovations could enhance MS4A6A detection in difficult sample types:

• Signal Amplification Technologies: Implement signal amplification methods such as tyramide signal amplification (TSA), rolling circle amplification (RCA), or proximity ligation assay (PLA) to boost detection sensitivity for low-abundance MS4A6A expression.

• Tissue Clearing Techniques: Combine MS4A6A antibodies with advanced tissue clearing methods (CLARITY, iDISCO, CUBIC) to enable whole-organ imaging with maintained protein antigenicity and improved antibody penetration.

• Multiplex Cyclic Immunofluorescence: Apply iterative staining and signal removal protocols to achieve highly multiplexed detection of MS4A6A alongside numerous other markers in the same tissue section.

• Automated Image Analysis: Develop machine learning algorithms specifically trained to recognize MS4A6A staining patterns in complex tissues, potentially overcoming limitations in visual assessment.

• Antigen Retrieval Optimization: Test emerging antigen retrieval technologies such as heat-mediated retrieval in specialized buffers or enzymatic digestion protocols optimized for membrane proteins like MS4A6A.

• Nanobody Development: Engineer MS4A6A-specific nanobodies that offer improved tissue penetration, reduced background, and greater epitope accessibility compared to conventional antibodies.

• Pre-embedding Immunolabeling: For electron microscopy applications, develop optimized pre-embedding immunolabeling protocols that preserve MS4A6A antigenicity while maintaining ultrastructural details.

• Sample-Specific Protocol Adaptation: Develop customized protocols for challenging sample types such as highly fibrotic tissues, lipid-rich environments, or samples with extensive post-mortem degradation.

How might experimental design evolve to address MS4A6A's complex biology in disease mechanisms?

Advanced experimental approaches will be necessary to fully elucidate MS4A6A's role in disease pathophysiology:

• Conditional Expression Systems: Design experiments using inducible expression or deletion of MS4A6A in specific cell types to dissect its function in complex disease models, particularly for neurodegenerative diseases and cancer.

• Domain-Specific Functional Analysis: Develop experimental designs using truncation mutants and targeted antibodies to investigate the function of specific MS4A6A domains in receptor complex formation and signal transduction.

• Multiscale Temporal Analysis: Implement experimental designs that capture MS4A6A dynamics across multiple timescales, from acute signaling events (seconds to minutes) to long-term expression changes during disease progression (months to years).

• Multimodal Data Integration: Design experiments that simultaneously capture MS4A6A protein expression, localization, interaction partners, and downstream signaling effects to build integrated models of its function.

• Systems Biology Approaches: Apply network analysis to position MS4A6A within broader signaling networks, identifying potential intervention points and feedback mechanisms.

• Cross-Disease Comparative Studies: Design parallel experiments across multiple disease models to identify common and disease-specific roles of MS4A6A, potentially revealing therapeutic opportunities with broader application.

• Humanized Models: Develop improved humanized models (e.g., patient-derived xenografts, organoids, or humanized mouse models) that better recapitulate human MS4A6A biology for more translational insights.

• Therapeutic Modulation: Design experiments to test antibody-based therapeutic approaches targeting MS4A6A, including blocking antibodies, antibody-drug conjugates, or bispecific antibodies linking MS4A6A-expressing cells to immune effectors.

What statistical approaches are recommended for analyzing quantitative MS4A6A expression data?

Appropriate statistical analysis is crucial for meaningful interpretation of MS4A6A expression data:

• Normalization Strategies: For Western blot or immunohistochemistry quantification, normalize MS4A6A signal to appropriate loading controls (e.g., β-actin, GAPDH) or reference proteins expressed in the same cell types.

• Statistical Tests Selection: For comparing MS4A6A expression between groups:

  • For normally distributed data: Apply parametric tests such as t-tests (two groups) or ANOVA (multiple groups)

  • For non-normally distributed data: Use non-parametric alternatives like Mann-Whitney U (two groups) or Kruskal-Wallis (multiple groups)

  • For paired samples: Select paired t-tests or Wilcoxon signed-rank tests as appropriate

• Multiple Testing Correction: When performing multiple comparisons, apply appropriate corrections (e.g., Bonferroni, Benjamini-Hochberg) to control false discovery rates.

• Correlation Analysis: For exploring relationships between MS4A6A expression and clinical parameters, use Pearson's correlation (linear, parametric) or Spearman's rank correlation (non-parametric) with appropriate visualization through scatter plots.

• Survival Analysis: For cancer studies, apply Kaplan-Meier analysis with log-rank tests to assess relationships between MS4A6A expression levels and patient outcomes, followed by multivariate Cox regression to adjust for confounding factors .

• Multivariate Analysis: Implement principal component analysis (PCA) or similar dimensionality reduction techniques to understand how MS4A6A expression relates to other molecular markers in complex datasets.

• Sample Size Considerations: Perform power analysis to determine appropriate sample sizes needed to detect biologically meaningful differences in MS4A6A expression with adequate statistical power.

• Reporting Standards: Adhere to best practices for statistical reporting, including clear documentation of statistical tests, exact p-values, confidence intervals, and effect sizes.

How should researchers interpret MS4A6A antibody signals in the context of heterogeneous tissue samples?

Interpreting MS4A6A staining in complex tissues requires careful consideration of several factors:

• Cellular Heterogeneity: Recognize that bulk tissue measurements may mask cell-type specific expression patterns. Use co-staining with cell-type markers (e.g., CD68, CD163 for macrophages) to attribute MS4A6A expression to specific cell populations .

• Spatial Context Analysis: Assess whether MS4A6A-expressing cells show particular spatial distributions within tissues (e.g., perivascular locations, tumor margins, inflammatory foci) that might suggest functional significance.

• Signal Intensity Quantification: Develop objective quantification methods that can distinguish between different levels of MS4A6A expression, potentially using digital image analysis with appropriate thresholding.

• Subcellular Localization: Evaluate whether MS4A6A staining shows expected membrane-associated patterns or unexpected subcellular distributions that might indicate altered trafficking or processing.

• Background Differentiation: Establish reliable methods to distinguish specific MS4A6A signal from background or non-specific staining, using appropriate negative controls and competing peptides where available.

• Cross-Validation: Validate observations using orthogonal methods (e.g., in situ hybridization for mRNA, mass spectrometry for protein) to confirm expression patterns observed with antibodies.

• Contextual Interpretation: Consider how disease state, treatment effects, or other experimental variables might influence MS4A6A expression patterns beyond simple presence/absence determinations.

• Technical Limitations Acknowledgment: Recognize and document limitations of antibody-based detection, including potential cross-reactivity, epitope masking, or differential sensitivity across sample types.

What are best practices for reporting MS4A6A antibody-based research findings in publications?

To ensure reproducibility and transparency in MS4A6A antibody-based research, adhere to these reporting standards:

• Antibody Documentation: Provide complete information about the MS4A6A antibody used, including:

  • Supplier and catalog number

  • Clone name for monoclonal antibodies

  • Host species and isotype

  • Immunogen details (e.g., the specific peptide sequence used)

  • Lot number when relevant for polyclonal antibodies

• Validation Evidence: Include explicit descriptions of antibody validation methods employed, such as:

  • Western blot data showing bands of expected molecular weight

  • Peptide competition assays

  • Positive and negative control tissues

  • Genetic validation approaches where applicable

• Protocol Transparency: Provide detailed methodological descriptions including:

  • Complete sample preparation procedures

  • Antigen retrieval methods and conditions

  • Antibody dilutions and incubation conditions

  • Detection systems and visualization methods

  • Image acquisition parameters

• Quantification Methods: Clearly describe any quantification approaches used, including:

  • Software tools and versions

  • Region of interest selection criteria

  • Thresholding parameters

  • Normalization strategies

• Representative Images: Include representative images that accurately reflect the full range of observations, not just the most dramatic examples. Provide scale bars and indicate magnification.

• Data Availability: Consider sharing original unprocessed images through appropriate repositories to enable re-analysis by other researchers.

• Limitations Acknowledgment: Explicitly discuss limitations of the antibody-based approaches used and how they might impact interpretation of results.

• Adherence to Guidelines: Follow established reporting guidelines such as ARRIVE for animal studies, REMARK for prognostic marker studies, or MIQE for qPCR studies when MS4A6A antibodies are used in these contexts.