S100A10 Antibody

What is S100A10 and why is it significant in research?

S100A10 (also known as p11) is an 11 kDa protein belonging to the S100 family of small, EF hand-containing dimeric proteins. It exists either as a monomer or as part of a heterotetrameric complex with Annexin A2 (ANXA2) . S100A10 has gained significant research attention due to its roles in:

• Cancer progression and development, particularly in hepatocellular carcinoma (HCC)

• Regulation of cellular processes including cell cycle progression

• Multidrug resistance in cancer cells

• Immune evasion mechanisms via CD8+ T cell exhaustion

• Potential as a biomarker for various cancer types

Recent studies have demonstrated that S100A10 is highly expressed in liver progenitor and premature hepatocyte stages compared to mature hepatocytes, with significant upregulation in HCC tumors compared to non-tumorous liver tissue .

What are the common applications for S100A10 antibodies?

S100A10 antibodies are utilized across multiple experimental platforms:

For optimal results, researchers should validate antibody performance in their specific experimental system and sample type.

How should I determine the appropriate dilution for S100A10 antibody applications?

Optimal dilution varies by application, antibody type, and sample source. Based on published data:

Always perform a titration experiment with your specific antibody and sample to determine optimal conditions. For example, when using rat anti-mouse S100A10 monoclonal antibody (MAB2377) for Western blot, 1 μg/mL successfully detected S100A10 in mouse lung tissue and MEF cells .

What protocols are most effective for detecting S100A10 in tissue samples?

For robust S100A10 detection in tissues, consider these methodological recommendations:

For Western Blot:

• Use PVDF membrane for optimal protein binding

• Apply reducing conditions with appropriate buffer systems (e.g., Immunoblot Buffer Group 1)

• Look for specific bands at approximately 11 kDa

• Include positive controls such as mouse lung tissue or embryonic feeder cells

For Immunohistochemistry:

• For optimal antigen retrieval, use TE buffer pH 9.0 (alternative: citrate buffer pH 6.0)

• When staining FFPE samples, test both buffer systems as retrieval efficacy can vary by tissue fixation

• Include appropriate negative controls using isotype-matched antibodies

• For colocalization studies, S100A10 is predominantly found in the cytoplasm

In a study using S100A10 antibody, researchers successfully detected cytoplasmic staining in TK-1 mouse T cell lymphoma lines at 5 μg/mL concentration, incubated for 3 hours at room temperature .

How can I optimize S100A10 detection in different cell types?

Cell type-specific optimization is critical for accurate S100A10 detection:

For adherent cells (e.g., MEF, HeLa, A431):

• For IF/ICC, grow cells on coverslips or chamber slides to 70-80% confluence

• Fix with 4% paraformaldehyde for 15-20 minutes at room temperature

• Permeabilize with 0.1-0.5% Triton X-100

• Block with 1-5% BSA or normal serum from the secondary antibody host species

• Incubate with primary antibody (e.g., 1:200-1:800 dilution) overnight at 4°C

For non-adherent cells (e.g., TK-1, Raw264):

• For flow cytometry, collect 1×10^6 cells and wash in PBS

• Fix and permeabilize using a commercial kit (e.g., FlowX FoxP3 Fixation & Permeabilization Buffer Kit)

• Stain with primary antibody followed by fluorophore-conjugated secondary antibodies

• Include isotype controls (e.g., Rat IgG2A for MAB2377)

For Raw264 mouse monocyte/macrophage cells, researchers successfully used permeabilization followed by staining with rat anti-mouse S100A10 monoclonal antibody and PE-conjugated secondary antibody .

How can S100A10 antibodies be used to study cancer progression mechanisms?

S100A10 antibodies have proven valuable for investigating cancer progression:

For studying HCC development:

• Use Western blot and IHC to assess S100A10 expression levels in tumor vs. non-tumor tissues

• Correlate expression with clinicopathological features (e.g., venous invasion, tumor differentiation)

• Implement S100A10 knockdown/knockout models to evaluate functional effects:

  • Xenograft tumor models show that S100A10 knockdown significantly reduces tumor volume

  • S100A10 overexpression increases chemoresistance to sorafenib, cisplatin, and 5-FU

For investigating metastasis:

• Use transwell migration/invasion assays with S100A10-overexpressing or knockdown cells

• Employ in vivo metastasis models:

  • Liver metastasis model: intrasplenic injection

  • Lung metastasis model: tail vein injection

Data from these models revealed that all mice injected with S100A10-overexpressing 97L cells developed metastatic liver nodules, while S100A10 knockout significantly inhibited metastasis .

What approaches can be used to study S100A10 in extracellular vesicles (EVs)?

S100A10 in EVs represents an emerging research area with specific methodological requirements:

• EV isolation protocols:

  • Differential ultracentrifugation

  • Size exclusion chromatography

  • Precipitation methods

• Verification of S100A10 in EVs:

  • Western blot analysis of EV lysates

  • Immunogold electron microscopy for localization in EVs

  • Flow cytometry of EV-bound beads

• Functional studies:

  • Co-culture recipient cells with S100A10-enriched EVs

  • Use S100A10-neutralizing antibodies to block effects

  • Assess downstream signaling (EGFR, AKT, ERK pathways)

Research has demonstrated that S100A10 is secreted by HCC cells into EVs both in vitro and in patient plasma. S100A10-enriched EVs enhance stemness and metastatic ability of recipient HCC cells and promote epithelial-mesenchymal transition. Importantly, S100A10 mediates the binding of MMP2, fibronectin, and EGF to EV membranes through interaction with integrin αV .

How can I investigate S100A10's role in immune modulation?

To study S100A10's impact on immune responses:

• For T cell exhaustion studies:

  • Co-culture CD8+ T cells with S100A10-expressing or silenced tumor cells

  • Analyze exhaustion markers (PD-1, TIM-3, LAG-3) by flow cytometry

  • Examine cytokine production (IFN-γ, IL-2)

  • Assess T cell proliferation and cytotoxicity

• For lipid metabolism pathway analysis:

  • Investigate cPLA2 and 5-LOX axis activation

  • Measure LTB4 levels in culture supernatants

  • Perform Co-IP experiments to identify protein-protein interactions

Recent research using HCC mouse models showed that S100A10 may activate the cPLA2 and 5-LOX axis, initiating lipid metabolism reprogramming and upregulating LTB4 levels, thereby promoting CD8+ T cell exhaustion and facilitating immune evasion by HCC cells .

How can I validate the specificity of S100A10 antibodies?

Robust validation ensures reliable experimental outcomes:

• Positive controls:

  • Known S100A10-expressing tissues (mouse lung, human lung)

  • Cell lines with confirmed expression (MEF, A431, HeLa)

• Negative controls:

  • Isotype-matched control antibodies (e.g., Rat IgG2A for MAB2377)

  • S100A10 knockout or knockdown samples

• Cross-reactivity assessment:

  • Test against related proteins (other S100 family members)

  • Note that some antibodies show partial cross-reactivity (e.g., ~5% cross-reactivity with recombinant human S100A10)

• Multiple detection methods:

  • Confirm results using different technical approaches (WB, IHC, IF)

  • Use antibodies targeting different epitopes

For example, when validating rat anti-mouse S100A10 monoclonal antibody, Western blot analysis confirmed a specific band at approximately 11 kDa in mouse lung tissue and MEF cells .

What are common challenges when working with S100A10 antibodies?

Researchers should be aware of these potential challenges:

For flow cytometry applications, proper cell fixation and permeabilization are critical for intracellular S100A10 detection. The FlowX FoxP3 Fixation & Permeabilization Buffer Kit has been successfully used for Raw264 mouse monocyte/macrophage cells .

How can S100A10 antibodies contribute to biomarker development?

S100A10's potential as a biomarker spans multiple clinical contexts:

Research has demonstrated that S100A10 protein in EVs serves as a potential biomarker for HCC detection and represents a promising therapeutic target .

What novel therapeutic approaches target S100A10?

Emerging therapeutic strategies involving S100A10 include:

• Neutralizing antibody approaches:

  • S100A10-neutralizing antibodies can block EV-mediated enhancement of cancer cell stemness and migration

  • This approach has shown efficacy in abrogating S100A10-mediated effects in experimental models

• Targeting S100A10-dependent pathways:

  • Interrupting S100A10's interaction with integrin αV

  • Inhibiting downstream signaling (EGFR, AKT, ERK pathways)

  • Modulating the cPLA2 and 5-LOX axis to prevent T cell exhaustion

• Combined approaches:

  • Using S100A10-targeting strategies alongside conventional treatments

  • Potential to overcome chemoresistance (sorafenib, cisplatin, 5-FU)

Recent research suggests that blockage of EV-S100A10 with S100A10-neutralizing antibody significantly abrogates enhancing effects on cancer progression, highlighting a promising therapeutic direction .

What are the recommended protocols for S100A10 antibody preparation?

For researchers developing their own S100A10 antibodies:

• Recombinant protein production:

  • Express full-length S100A10 in E. coli BL21(DE3) using pET28a(+) vector

  • Induce with IPTG and purify using Ni-NTA resin

• Polyclonal antibody production:

  • Use purified recombinant S100A10 protein as antigen

  • Inoculate rabbits intradermally

  • Assess antibody titer by ELISA

  • Evaluate specificity by Western blotting

One study successfully established a prokaryotic expression and purification system for S100A10 and generated polyclonal antibodies with high titer and specificity, providing valuable tools for further S100A10 research .