SH2D3C Antibody

What is SH2D3C and what are its primary functions in cellular biology?

SH2D3C (SH2 domain-containing protein 3C), also known as NSP3, is an adaptor protein belonging to a cytoplasmic protein family primarily involved in cell migration. This protein contains both an SH2 domain and a guanine nucleotide exchange factor-like domain (Ras GEF-like domain) that allows it to form complexes with scaffolding proteins such as Crk-associated substrate . SH2D3C functions as a signaling molecule involved in multiple cellular processes including adhesion, migration, tissue organization, and immune response regulation . It has been identified as an Eph receptor-binding protein that may positively regulate T-cell receptor (TCR) signaling . The protein exists in multiple isoforms with calculated molecular weights ranging from 77-94 kDa, though the observed molecular weight in most applications is approximately 77 kDa .

Which applications are SH2D3C antibodies validated for?

SH2D3C antibodies have been validated for multiple research applications with specific recommended parameters:

It is recommended to titrate these antibodies in each testing system to obtain optimal results, as sensitivity may be sample-dependent .

How should SH2D3C antibodies be stored and handled to maintain reactivity?

For optimal maintenance of antibody reactivity, SH2D3C antibodies should be stored at -20°C where they remain stable for approximately one year after shipment. Many commercial preparations are supplied in PBS with 0.02% sodium azide and 50% glycerol at pH 7.3 . For lyophilized antibody formulations, reconstitution should be performed with sterile deionized water to achieve the recommended concentration (typically 0.5 mg/ml) .

After reconstitution, the antibody can be stored at 4°C for up to one month, but for long-term storage, it should be aliquoted and kept at -20°C to avoid repeated freeze-thaw cycles that can diminish activity . Some smaller antibody preparations (e.g., 20 μl sizes) may contain 0.1% BSA as a stabilizer . Always centrifuge the antibody vial before opening to ensure collection of the entire solution.

What are the critical considerations for successful Western blot detection of SH2D3C?

When designing Western blot experiments for SH2D3C detection, researchers should consider several critical factors:

• Sample selection: Human, mouse, and rat samples have demonstrated reactivity with common SH2D3C antibodies. HEK-293 cells, human brain tissue, and human placenta tissue have been validated as positive controls .

• Molecular weight considerations: Multiple SH2D3C isoforms exist with calculated molecular weights of 94 kDa, 86 kDa, and 77 kDa, though the predominantly observed band appears at approximately 77 kDa. When working with human placental tissue lysate, predicted molecular weights range from 55-94 kDa due to multiple isoforms .

• Antibody dilution: A titration range of 1:500-1:2400 or 1-2 μg/ml is recommended, with optimization needed for each specific sample type .

• Detection system: Secondary antibody selection should be compatible with the primary antibody host (typically rabbit IgG for many commercial SH2D3C antibodies) .

• Negative controls: Include samples known to lack SH2D3C expression or use isotype controls to verify specificity.

A methodical approach to optimization involves testing multiple antibody concentrations against positive control samples, followed by application of optimized conditions to experimental samples.

How can immunoprecipitation protocols be optimized for studying SH2D3C interactions?

Optimization of immunoprecipitation (IP) protocols for SH2D3C studies requires careful consideration of several parameters, particularly when investigating protein-protein interactions:

• Antibody amount: Use 0.5-4.0 μg of SH2D3C antibody per 1.0-3.0 mg of total protein lysate. This ratio may need adjustment based on expression levels in your sample .

• Validated samples: Mouse brain tissue has been confirmed as suitable for SH2D3C immunoprecipitation .

• Protocol modifications for interaction studies: When investigating protein interactions such as the DPP3–SH2D3C interaction, consider these approaches:

  • For overexpression studies, FLAG-tagged SH2D3C isoforms can be transiently overexpressed in cells (e.g., TRex HEK293T) stably expressing the protein of interest with an alternative tag (e.g., HA-tag) .

  • Use agarose-conjugated tag-specific antibodies (e.g., anti-HA) for co-immunoprecipitation followed by Western blot analysis with antibodies against the interacting protein's tag (e.g., anti-FLAG) .

  • Include appropriate controls: negative controls (cells transfected with empty vector) and positive controls (known interacting proteins) .

• Buffer considerations: Gentle lysis buffers that preserve protein-protein interactions while efficiently extracting membrane-associated proteins are recommended.

The DPP3–SH2D3C interaction was successfully demonstrated using this approach, confirming binding between DPP3 and both isoforms 2 and 3 of SH2D3C protein .

What are the recommended antigen retrieval methods for IHC detection of SH2D3C?

For optimal immunohistochemical detection of SH2D3C in tissue samples, antigen retrieval methodology significantly impacts staining quality and specificity. The recommended protocols are:

• Primary recommendation: Antigen retrieval with TE buffer at pH 9.0. This alkaline pH helps break protein cross-links formed during fixation and unmasks antigenic sites, particularly effective for SH2D3C detection in human breast cancer tissue .

• Alternative method: Citrate buffer at pH 6.0 can be used as an alternative approach if the primary method yields suboptimal results .

• Antibody dilution: A wide range of 1:20-1:200 is recommended for IHC applications, indicating that optimization is required for each tissue type and experimental condition .

• Controls: Include positive control tissues (human breast cancer tissue has been validated) and negative controls (omission of primary antibody or use of isotype control) .

The need for two different pH environments for antigen retrieval suggests that SH2D3C epitopes may be differentially affected by fixation methods, and researchers should systematically compare both approaches when establishing protocols for new tissue types.

How can SH2D3C antibodies be utilized to investigate its role in cancer progression?

SH2D3C has emerged as a significant factor in cancer biology, particularly in lung cancer where it's associated with advanced stage and poor prognosis . Researchers investigating its role in cancer progression can implement these methodological approaches:

• Prognostic biomarker studies:

  • Use IHC with SH2D3C antibodies (1:20-1:200 dilution) on tumor tissue microarrays to correlate expression levels with clinical outcomes .

  • Complement tissue studies with serum analysis using ELISA (0.1-0.5 μg/ml antibody concentration) to evaluate potential as a circulating biomarker .

• Tumor immune microenvironment analysis:

  • Employ flow cytometry (1-3 μg/million cells) on tumor-infiltrating lymphocytes to investigate SH2D3C's role in T-cell dysfunction and exclusion mechanisms .

  • Combine with antibodies against T-cell markers to assess correlation between SH2D3C expression and immune cell phenotypes.

• Genetic alteration correlation:

  • When studying lung cancer specifically, analyze the inverse relationship between SH2D3C alterations and EGFR alterations through combined IHC and genetic analysis .

  • Use Western blotting (1:500-1:2400) to assess protein expression changes in relation to genetic status .

• Therapy resistance mechanisms:

  • Employ SH2D3C antibodies in post-treatment tumor samples to evaluate expression changes in therapy-resistant versus responsive tumors .

  • Develop in vitro models with varied SH2D3C expression to assess drug sensitivity using flow cytometry and Western blot analysis .

These approaches can provide insights into how SH2D3C contributes to cancer progression and therapy resistance, potentially identifying new therapeutic strategies targeting this protein.

What approaches can be used to study the interaction between SH2D3C and DPP3 in oxidative stress regulation?

The recently identified interaction between SH2D3C and Dipeptidyl peptidase 3 (DPP3) represents a potentially important link between oxidative stress response and cellular migration pathways . Researchers can employ these methodological approaches to investigate this interaction:

• Co-immunoprecipitation validation:

  • Perform bidirectional co-IPs using both SH2D3C (0.5-4.0 μg for 1.0-3.0 mg lysate) and DPP3 antibodies on endogenous proteins in relevant cell types .

  • For cells with low SH2D3C expression, consider targeted enhancement using CRISPR activation systems rather than overexpression to maintain physiological relevance.

• Domain mapping studies:

  • Generate constructs expressing specific domains of SH2D3C (particularly the SH2 domain and Ras GEF-like domain) to determine which regions are essential for DPP3 interaction.

  • Use IP with domain-specific antibodies followed by Western blot analysis to map the interaction interface .

• Functional impact on KEAP1-NRF2 pathway:

  • Employ SH2D3C antibodies (1:500-1:2400 dilution) in Western blots to monitor changes in KEAP1-NRF2 pathway components when SH2D3C levels are manipulated .

  • Use oxidative stress inducers to test how the SH2D3C-DPP3 interaction affects cellular responses to stress conditions.

• Subcellular localization:

  • Utilize IHC or immunofluorescence (1:20-1:200 dilution) to investigate colocalization of SH2D3C and DPP3 under different cellular conditions .

  • Perform fractionation studies followed by Western blot analysis to determine compartment-specific interactions.

This systematic approach can help elucidate how SH2D3C-DPP3 interaction contributes to the regulation of oxidative stress response through the KEAP1–NRF2 pathway, potentially revealing new therapeutic targets for diseases involving oxidative stress.

How can flow cytometry with SH2D3C antibodies be optimized for analyzing immune cell populations?

Flow cytometry using SH2D3C antibodies can provide valuable insights into immune cell phenotypes and functions, particularly given SH2D3C's role in T-cell trafficking and immune regulation . Optimal protocols should consider:

• Cell preparation and fixation:

  • Use fixation and permeabilization for intracellular SH2D3C detection, as validated in human HEL cells .

  • Optimize fixation time and permeabilization buffer composition based on cell type to maximize signal while preserving cellular integrity.

• Antibody concentration and blocking:

  • Use 1-3 μg SH2D3C antibody per million cells as a starting point .

  • Block with appropriate sera (goat sera has been validated) to minimize non-specific binding .

• Multi-parameter panel design:

  • Include markers for specific immune cell populations (e.g., CD3, CD4, CD8 for T-cells) alongside SH2D3C to correlate expression with cellular phenotypes.

  • Consider activation markers (CD69, CD25) to assess relationship between SH2D3C expression and immune cell activation status.

• Controls and gating strategy:

  • Include cells alone (unstained), isotype controls, and single-stained controls as demonstrated in validated protocols .

  • Implement a hierarchical gating strategy starting with intact cells → single cells → specific immune populations → SH2D3C expression.

• Signal calibration:

  • Use quantitative beads to standardize fluorescence intensity and enable comparison across experiments and between different immune cell subsets.

This methodological approach enables robust analysis of SH2D3C expression in diverse immune cell populations, facilitating investigation of its role in normal immune function and dysfunction in disease states.

What are common issues in SH2D3C antibody applications and how can they be resolved?

When working with SH2D3C antibodies, researchers may encounter several technical challenges that require systematic troubleshooting:

• Western blot detection issues:

  • Problem: Multiple bands or unexpected molecular weights.
    Solution: SH2D3C has multiple isoforms (calculated MW: 94, 86, and 77 kDa), with 77 kDa being the predominantly observed band. Ensure appropriate positive controls (HEK-293 cells, human brain tissue, or human placenta tissue) to confirm banding patterns .

  • Problem: Weak or no signal.
    Solution: Increase antibody concentration within the recommended range (1:500-1:2400), extend incubation time, or try enhanced chemiluminescence detection systems .

• Immunoprecipitation challenges:

  • Problem: Low yield of immunoprecipitated protein.
    Solution: Increase antibody amount up to 4.0 μg per 1.0-3.0 mg of total protein lysate. Consider using agarose-conjugated antibodies for more efficient capture .

  • Problem: Non-specific binding in co-IP experiments.
    Solution: Implement more stringent washing conditions and include appropriate blocking proteins in lysis and wash buffers .

• Immunohistochemistry inconsistencies:

  • Problem: Variable staining intensity or background.
    Solution: Optimize antigen retrieval by comparing TE buffer (pH 9.0) versus citrate buffer (pH 6.0). Adjust antibody dilution within the 1:20-1:200 range for optimal signal-to-noise ratio .

• Flow cytometry issues:

  • Problem: Poor discrimination between positive and negative populations.
    Solution: Ensure proper fixation and permeabilization for intracellular staining. Use 1-3 μg antibody per million cells with appropriate blocking sera (e.g., goat sera) as validated in protocols .

• Cross-reactivity concerns:

  • Problem: Potential antibody cross-reactivity with related proteins.
    Solution: Validate antibody specificity using known positive controls (human, mouse, rat samples have demonstrated reactivity) and negative controls (isotype antibodies or samples with SH2D3C knockdown) .

Systematic optimization and thorough controls are essential for generating reliable data with SH2D3C antibodies across all applications.

How can researchers validate SH2D3C antibody specificity in their experimental systems?

Validating antibody specificity is crucial for generating reliable scientific data. For SH2D3C antibodies, researchers should implement these methodological approaches:

• Genetic validation techniques:

  • Perform siRNA/shRNA knockdown or CRISPR/Cas9 knockout of SH2D3C in relevant cell types, followed by Western blotting to confirm reduced or absent signal compared to control cells.

  • Alternatively, overexpress tagged SH2D3C (as demonstrated with FLAG-tagged isoforms 2 and 3) and confirm antibody detection of the overexpressed protein .

• Multiple antibody validation:

  • Compare results using antibodies from different sources or those targeting different epitopes of SH2D3C.

  • Confirm consistent patterns across antibodies, particularly regarding molecular weight detection (predicted bands at 55-94 kDa due to multiple isoforms) .

• Cross-reactivity assessment:

  • Test antibodies on tissues from multiple species to confirm expected reactivity patterns (validated in human, mouse, and rat samples) .

  • Include tissues known to express high levels of SH2D3C (brain, placenta) as positive controls .

• Application-specific controls:

  • For Western blot: Include recombinant SH2D3C protein as a positive control.

  • For IP: Perform reverse IP where possible and confirm results.

  • For IHC: Include isotype controls and tissues with known expression patterns.

  • For flow cytometry: Compare with isotype controls and cells alone (unstained) as demonstrated in validated protocols .

• Isoform-specific validation:

  • Design experiments to distinguish between SH2D3C isoforms, particularly when studying specific functions of isoforms 2 and 3 that have been shown to interact with proteins like DPP3 .

These validation approaches ensure that observed signals truly represent SH2D3C protein and provide confidence in subsequent experimental findings.

How can SH2D3C antibodies be utilized in neurodegeneration research?

Recent findings linking SH2D3C to neurodegeneration offer promising research directions. SH2D3C was found to be upregulated in rat cortical neurons after treatment with amyloid-β-oligomers, with higher protein levels detected in Alzheimer's disease mice compared to wild type . Researchers can employ these methodological approaches:

• Comparative expression analysis:

  • Use Western blotting (1:500-1:2400 dilution) to quantitatively compare SH2D3C levels in brain tissues from neurodegenerative disease models versus controls .

  • Apply IHC (1:20-1:200 dilution) to map regional distribution changes in the brain during disease progression .

• Mechanistic studies of neuronal death:

  • Since SH2D3C overexpression has been linked to neuronal death, use antibodies to monitor expression changes during neurodegeneration progression .

  • Combine with markers of neuronal viability and apoptosis in multi-parameter flow cytometry (1-3 μg/million cells) or immunofluorescence studies .

• Amyloid-β interaction studies:

  • Investigate whether SH2D3C directly interacts with amyloid-β using co-immunoprecipitation (0.5-4.0 μg antibody per 1.0-3.0 mg lysate) .

  • Correlate SH2D3C expression changes with amyloid plaque distribution using dual-label IHC techniques.

• Intervention assessment:

  • Apply SH2D3C antibodies to evaluate whether therapeutic interventions that modulate SH2D3C levels or function can mitigate neurodegeneration.

  • Monitor SH2D3C expression changes in response to neuroprotective compounds.

• Clinical correlation studies:

  • Develop protocols for analyzing SH2D3C in human cerebrospinal fluid or brain tissue samples from neurodegenerative disease patients.

  • Correlate expression levels with disease severity and progression using validated antibodies.

These approaches can help elucidate the role of SH2D3C in neurodegeneration, potentially identifying new therapeutic targets for Alzheimer's disease and related conditions.

What strategies can be employed to explore SH2D3C as a therapeutic target in cancer?

SH2D3C has emerged as a potential therapeutic target in cancer, particularly in lung cancer where it is associated with advanced stage, poor prognosis, and therapy resistance . Researchers can employ these methodological approaches:

• Target validation studies:

  • Use SH2D3C antibodies in IHC (1:20-1:200) and Western blot (1:500-1:2400) to correlate expression with clinical outcomes across cancer types .

  • Perform knockdown/knockout studies followed by functional assays to confirm SH2D3C's role in cancer cell survival, migration, and therapy resistance.

• Molecular docking and inhibitor development:

  • Building on existing research with organosulfur compounds from Allium sativum (garlic), develop screening assays to identify SH2D3C inhibitors .

  • Use antibodies to evaluate whether candidate compounds modulate SH2D3C expression or downstream signaling pathways.

• Combination therapy approaches:

  • Since SH2D3C alterations co-occur inversely with EGFR alterations in lung cancer, investigate combinations of SH2D3C-targeting compounds with existing EGFR inhibitors .

  • Use antibodies to monitor pathway activity in response to combination treatments.

• Immune checkpoint therapy correlation:

  • Given SH2D3C's role in promoting tumor immune evasion through T-cell dysfunction, use flow cytometry (1-3 μg/million cells) to investigate potential synergy between SH2D3C inhibition and immune checkpoint blockade .

  • Apply multiplexed IHC to evaluate changes in tumor immune microenvironment following SH2D3C modulation.

• Biomarker development:

  • Establish standardized IHC or ELISA protocols using validated antibodies to evaluate SH2D3C as a predictive biomarker for therapy selection .

  • Correlate expression levels with response to targeted therapies and immunotherapies.

This systematic approach can accelerate the development of SH2D3C-targeted therapeutics and identify patient populations most likely to benefit from such interventions.

How can SH2D3C expression be evaluated in challenging tissue samples?

Some tissue types present unique challenges for protein detection. When working with difficult samples for SH2D3C analysis, researchers should consider these methodological approaches:

• Optimization for formalin-fixed paraffin-embedded (FFPE) tissues:

  • Compare antigen retrieval methods: TE buffer (pH 9.0) is recommended as primary approach, with citrate buffer (pH 6.0) as an alternative .

  • Extend antigen retrieval time for highly fixed tissues while monitoring tissue integrity.

  • For breast cancer tissue specifically, optimize antibody dilution within the 1:20-1:200 range based on fixation variables .

• Fresh frozen tissue processing:

  • Modify fixation protocols for immunofluorescence to preserve SH2D3C epitopes while maintaining cellular architecture.

  • For Western blot analysis, optimize extraction buffers to efficiently solubilize SH2D3C from different tissue types (validated in brain and placenta tissues) .

• Brain tissue-specific considerations:

  • Given SH2D3C's expression in brain and its potential role in neurodegeneration, employ region-specific sampling to account for heterogeneous expression .

  • For mouse brain tissue IP, optimize lysis conditions to preserve protein-protein interactions while efficiently extracting SH2D3C .

• Tumor heterogeneity assessment:

  • Implement tissue microarray approaches with multiple cores per tumor to account for expression heterogeneity.

  • Combine with digital pathology quantification for objective assessment of expression patterns.

• Low-expression sample analysis:

  • Employ signal amplification systems (tyramide signal amplification for IHC or high-sensitivity ECL for Western blot) when working with tissues with low SH2D3C expression.

  • Consider enrichment techniques such as immunoprecipitation before Western blot analysis.

These approaches enable reliable detection of SH2D3C across diverse and challenging tissue types, expanding the range of samples suitable for analysis.

What considerations should be made when designing multiplexed assays involving SH2D3C antibodies?

Multiplexed assays provide comprehensive insights into protein interactions and pathway activities. When incorporating SH2D3C antibodies into multiplexed experimental designs, researchers should consider:

• Antibody compatibility assessment:

  • Test for cross-reactivity between SH2D3C antibodies and other antibodies in the multiplex panel.

  • For immunofluorescence or flow cytometry, select SH2D3C antibodies with compatible host species to avoid secondary antibody cross-reactivity .

• Flow cytometry panel design:

  • When designing multi-parameter flow cytometry panels (using 1-3 μg SH2D3C antibody per million cells), consider fluorophore brightness relative to SH2D3C expression level in target cells .

  • Include compensation controls to account for spectral overlap between fluorophores.

  • Validated in human HEL cells, but requires optimization for immune cell subpopulations .

• Multiplexed IHC/IF approaches:

  • For sequential immunostaining, determine optimal antibody stripping or quenching protocols that preserve tissue integrity while removing previous antibody layers.

  • For simultaneous staining, select antibodies raised in different host species and validate staining patterns individually before combining.

  • Test antibody performance at 1:20-1:200 dilution range in multiplex context, as optimal concentration may differ from single-stain applications .

• Protein array applications:

  • Validate SH2D3C antibody specificity in protein array format before incorporation into multiplexed arrays.

  • Establish signal-to-noise thresholds specific to array-based detection systems.

• Mass cytometry considerations:

  • For CyTOF applications, validate metal-conjugated SH2D3C antibodies for specificity and sensitivity compared to fluorophore-conjugated versions.

  • Optimize staining protocol for metal-labeled antibodies, which may differ from flow cytometry protocols.

These considerations enable reliable incorporation of SH2D3C detection into complex multiplexed assays, facilitating comprehensive analysis of its interactions and functions within cellular pathways.

How might new technologies enhance SH2D3C antibody applications in research?

Emerging technologies offer opportunities to extend the capabilities of SH2D3C antibody applications in several key areas:

• Single-cell analysis techniques:

  • Apply SH2D3C antibodies in single-cell Western blotting to assess expression heterogeneity within populations.

  • Develop protocols for compatible antibody panels for single-cell proteomic analysis using technologies like CITE-seq to correlate SH2D3C protein expression with transcriptomic profiles.

• Advanced imaging applications:

  • Adapt SH2D3C antibodies (1:20-1:200 dilution range) for super-resolution microscopy to map precise subcellular localization .

  • Implement live-cell imaging using cell-permeable fluorescently-labeled antibody fragments to track SH2D3C dynamics in real-time.

  • Develop protocols for expansion microscopy to visualize SH2D3C in the context of nanoscale cellular structures.

• Proximity labeling approaches:

  • Couple SH2D3C antibodies with proximity labeling enzymes (BioID, APEX) to identify novel protein interactions in living cells.

  • This could expand upon the recently discovered DPP3-SH2D3C interaction and identify additional partners in different cellular contexts .

• Antibody engineering innovations:

  • Develop recombinant antibody formats with enhanced tissue penetration for improved IHC applications.

  • Engineer bi-specific antibodies to simultaneously detect SH2D3C and its interaction partners like DPP3 .

• Clinical translation technologies:

  • Adapt validated SH2D3C antibodies for use in circulating tumor cell detection platforms.

  • Develop companion diagnostic assays based on standardized IHC protocols to identify patients with SH2D3C-driven cancers who might benefit from targeted therapies .

These technological advances would significantly enhance the utility of SH2D3C antibodies across basic research, translational studies, and potential clinical applications.

What are the most promising intersections between SH2D3C research and emerging therapeutic approaches?

Current research highlights several promising intersections between SH2D3C biology and emerging therapeutic approaches:

• Immuno-oncology applications:

  • Given SH2D3C's role in promoting tumor immune evasion through T-cell dysfunction and exclusion mechanisms, investigate combination approaches with immune checkpoint inhibitors .

  • Develop protocols using flow cytometry (1-3 μg antibody/million cells) to monitor how modulating SH2D3C affects immune cell composition in the tumor microenvironment .

• Targeted therapy development:

  • Building on molecular docking studies with organosulfur compounds from Allium sativum, extend to larger compound libraries to identify potent and selective SH2D3C inhibitors .

  • Use SH2D3C antibodies in Western blot (1:500-1:2400) and IP applications to assess compound effects on expression and pathway activation .

• Neurodegeneration intervention strategies:

  • Given the association between SH2D3C upregulation and neuronal death in Alzheimer's disease models, explore neuroprotective approaches targeting SH2D3C .

  • Apply IHC (1:20-1:200) to evaluate how candidate compounds affect SH2D3C expression in brain tissue .

• Oxidative stress modulation:

  • Investigate the therapeutic potential of modulating the DPP3-SH2D3C interaction to influence oxidative stress responses via the KEAP1-NRF2 pathway .

  • Employ co-IP approaches (0.5-4.0 μg antibody per 1.0-3.0 mg lysate) to screen for compounds that selectively disrupt or enhance this interaction .

• Biomarker-guided precision medicine:

  • Develop standardized IHC protocols with validated antibodies for patient stratification in clinical trials.

  • Particularly in lung cancer, where SH2D3C is associated with advanced stage and poor prognosis, correlate expression with response to targeted therapies and immunotherapies .

These intersections represent high-priority areas for translational research that could leverage SH2D3C biology for therapeutic benefit across multiple disease contexts.

How do different commercial SH2D3C antibodies compare in performance across applications?

When selecting an SH2D3C antibody for specific research applications, understanding performance differences between commercial options is essential:

Key comparative insights for application-specific selection:

• Western blot applications:

  • Both antibodies detect multiple isoforms, with Proteintech reporting predominant 77 kDa band while NSJ Bioreagents reports a broader 55-94 kDa range .

  • Consider the specific isoforms of interest when selecting an antibody.

• Immunoprecipitation strengths:

  • Proteintech antibody has validated protocols for IP applications in mouse brain tissue .

  • NSJ antibody lacks specific IP validation in the available data .

• Flow cytometry capabilities:

  • NSJ antibody has specific validation for flow cytometry in human HEL cells .

  • Proteintech antibody lacks specific flow cytometry validation in the available data .

• Storage and handling differences:

  • Proteintech antibody is supplied in liquid form with glycerol .

  • NSJ antibody is lyophilized and requires reconstitution .

When selecting an antibody, researchers should prioritize those with validation in their specific application and tissue/cell type of interest, and conduct comparative testing when possible to determine optimal performance in their experimental system.

What are the key differences between studying SH2D3C in cancer versus neurodegeneration contexts?

SH2D3C plays distinct roles in cancer and neurodegeneration, necessitating different methodological approaches and considerations:

Research design considerations for each context:

• Cancer-specific approaches:

  • Prioritize antibody applications in diverse tumor types and immune cell populations .

  • Focus on correlation with clinical outcomes and therapy response.

  • Investigate relationship with immune checkpoint molecules and T-cell function.

• Neurodegeneration-specific approaches:

  • Optimize antibody protocols for brain tissue specificity and regional analysis .

  • Emphasize co-localization studies with amyloid-β and other neurodegeneration markers.

  • Investigate temporal expression changes during disease progression.

These distinct approaches reflect the different cellular contexts and pathological mechanisms through which SH2D3C contributes to cancer and neurodegeneration, requiring carefully tailored experimental designs.