Cell Biology Research Tools: CD14+ Cell Sorting & PKCζ-Mediated ADPKD Mechanisms

Concept

CD14-positive monocytes/macrophages are central effector cells in immunology, inflammation and disease pathogenesis, while Protein Kinase C zeta (PKCζ) is a master regulator of renal epithelial cell function and a key molecular target in Autosomal Dominant Polycystic Kidney Disease (ADPKD)—two critical research areas in modern cell and disease biology. As a leading global provider of life science reagents, ANT BIO PTE. LTD. delivers cutting-edge research tools for both fields under its Starter sub-brand (our flagship line for high-performance antibodies and cell isolation kits): the Starter MagSep™ CD14 Positive Cell Sorting Kit for rapid, high-purity isolation of human CD14+ monocytes, and the PKCζ Recombinant Rabbit Monoclonal Antibody for dissecting PKCζ-mediated signaling mechanisms in ADPKD and beyond. These tools combine technical precision, ease of use and broad application utility, empowering breakthroughs in immunology, nephrology, cell signaling and translational drug development.

Research Frontiers

The CD14+ cell sorting kits and PKCζ recombinant rabbit monoclonal antibody from ANT BIO PTE. LTD. enable cutting-edge research across immunology, nephrology, cell biology and translational science—fields where high-purity cell isolation and isoform-specific signal transduction analysis are foundational. Key research frontiers unlocked by these tools include:

1. CD14+ monocyte/macrophage immunology: Dissecting the role of CD14+ cells in innate immunity, inflammation, autoimmune disease, tumor microenvironment and atherosclerosis.
2. ADPKD pathogenesis and therapy: Elucidating PKCζ-mediated polycystin-1 phosphorylation and its regulatory role in renal cyst formation, epithelial polarity and ciliogenesis in inherited kidney disease.
3. Cell signaling cross-talk: Exploring intersections between CD14/TLR4 innate immune signaling and PKCζ-mediated cell polarity pathways in disease.
4. Cell therapy and regenerative medicine: Using high-purity CD14+ monocytes as precursor cells for macrophage/dendritic cell differentiation and cell-based immunotherapies.
5. Repurposed drug development: Validating PKCζ activators (e.g., fingolimod) for ADPKD and CD14-targeted therapies for inflammatory and autoimmune diseases.
6. Single-cell and omics research: Providing high-purity CD14+ cell populations for single-cell sequencing, transcriptomics and proteomics, and PKCζ tools for molecular mechanism analysis in these high-throughput studies.
7. Disease model validation: Using CD14+ sorting kits to construct immune inflammation models and PKCζ antibodies to validate ADPKD preclinical models for therapeutic testing.

Research Significance

CD14+ monocytes/macrophages and PKCζ-mediated signaling represent two critical pillars of disease biology, with profound implications for understanding and treating human disease. ANT BIO PTE. LTD.’s specialized research tools amplify the scientific and translational significance of these areas by delivering the precision, reliability and efficiency needed for modern research:

1. Enabling high-purity immune cell research: The CD14+ sorting kit provides a fast, user-friendly method to isolate high-viability CD14+ monocytes—eliminating the need for complex flow cytometry equipment and making pure innate immune cell populations accessible to all laboratories, accelerating immunology research.
2. Unlocking ADPKD molecular mechanisms: The PKCζ antibody enables isoform-specific analysis of PKCζ function in ADPKD, defining the PKCζ-polycystin-1 axis as a novel therapeutic target for the most common inherited kidney disease (affecting 12+ million worldwide).
3. Bridging cell isolation and molecular analysis: Together, these tools create a seamless workflow for disease research—from isolating disease-relevant immune/renal cells to dissecting the molecular signaling pathways that drive their dysfunction.
4. Accelerating translational research: The CD14+ kit supports the development of cell-based therapies and inflammatory disease biomarkers, while the PKCζ antibody validates novel therapeutic targets for ADPKD, a disease with no curative treatments.
5. Increasing research accessibility: The MagSep™ CD14 kit’s simple operation and low equipment requirements democratize high-purity cell sorting, while the PKCζ antibody’s multi-platform utility (WB, IP, IF, IHC) and pan-species reactivity (human/mouse/rat) make it adaptable to all preclinical and translational research models.
6. Ensuring experimental reproducibility: Rigorously validated, batch-consistent tools standardize experimental workflows across laboratories, improving data comparability and reproducibility in both CD14+ immunology and PKCζ signaling research.

Related Mechanisms, Research Methods & Product Applications

1. CD14+ Cell Sorting: Key Technologies, Optimization and Starter MagSep™ Kit Applications

1.1 Why CD14+ Cell Isolation Is Critical for Immunology Research

CD14 is a glycosylphosphatidylinositol-anchored protein expressed on the surface of monocytes, macrophages and myeloid dendritic cells, serving as a co-receptor for Toll-like receptor 4 (TLR4) and a key marker for innate immune cell identification. CD14+ monocytes are the primary precursors of tissue macrophages and dendritic cells, and their dysfunction drives the pathogenesis of numerous diseases—including inflammation, autoimmune disorders, atherosclerosis and cancer. Isolating high-purity, high-viability CD14+ cells is essential for studying their phenotypic and functional characteristics, and for developing cell-based therapies targeting innate immune dysfunction.

1.2 Key Considerations for CD14+ Cell Sorting Sample Preparation

Successful CD14+ cell isolation begins with rigorous sample preparation, the foundation of high-purity, high-viability cell recovery:

1. Cell Source and Viability: Use fresh peripheral blood mononuclear cells (PBMCs), whole blood, leukocyte-enriched products or tissue cell suspensions; ensure cell viability >90% and intact surface CD14 antigen expression before sorting.
2. Antibody Labeling Optimization: Select highly specific anti-CD14 antibodies with minimal non-specific binding; optimize antibody dosage (1/4–1/5 of conventional staining volumes) to reduce costs while maintaining labeling efficiency.
3. Cell Concentration Control: Adjust cell concentration to 10⁶–10⁷ cells/mL to avoid aggregation and nozzle clogging (for flow sorting) or inefficient magnetic separation.
4. Buffer Selection: Use phosphate-buffered saline (PBS) or Hank’s Balanced Salt Solution (HBSS) containing 2–5% fetal bovine serum (FBS) or bovine serum albumin (BSA) as staining/separation buffer to reduce non-specific binding and maintain cell viability.

1.3 Optimization Strategies for CD14+ Cell Sorting Recovery and Viability

Whether using flow cytometry or magnetic bead sorting, systematic optimization is critical to maximize CD14+ cell recovery, purity and viability:

1. Pre-Sorting Treatment: Use siliconized/low-adsorption polypropylene consumables to minimize cell loss via adhesion; perform accurate cell counting for baseline recovery rate calculations.
2. Instrument/Device Parameter Optimization: For flow sorting, optimize nozzle diameter (70–100 μm), sheath fluid (HBSS preferred over PBS) and deflection voltage; for magnetic sorting, use optimized magnetic separation stands for efficient bead-cell complex capture.
3. Sorting Mode Selection: Use enrichment mode for high-abundance CD14+ cells to increase speed; use purity mode for rare cells or ultra-high purity requirements (≥95%).
4. Collection Condition Control: Pre-fill collection tubes with culture medium containing 20–30% FBS; gently invert tubes during sorting to prevent sedimentation; centrifuge sorted cells at 300–400g within 10 minutes to avoid viability loss.
5. Temperature Control: Perform sorting at 4°C, 15°C or room temperature based on cell type; cold conditions minimize metabolic stress and preserve surface antigen expression.

1.4 Quality Evaluation of Sorted CD14+ Cells

Comprehensive post-sorting quality assessment is essential to ensure CD14+ cells are suitable for downstream experiments:

1. Purity Verification: Re-stain sorted cells with anti-CD14 antibodies and analyze via flow cytometry; target purity ≥90% (magnetic sorting) or ≥95% (flow sorting).
2. Viability Detection: Use trypan blue exclusion or live/dead fluorescent staining; target post-sorting viability ≥85%.
3. Functional Status Assessment: Evaluate phagocytic activity, cytokine secretion (e.g., TNF-α, IL-6) or differentiation potential (M1/M2 macrophages) to confirm functional integrity.
4. Sterility and Counting: Perform sterility testing for cell culture experiments; precisely count sorted cells to calculate recovery rate (actual yield/expected yield × 100%).

1.5 Starter MagSep™ CD14 Positive Cell Sorting Kit (S0D3008): Core Advantages and Applications

ANT BIO PTE. LTD.’s Starter MagSep™ CD14 Positive Cell Sorting Kit (S Type, S0D3008) is a magnetic bead-based cell isolation system engineered for fast, high-purity isolation of human CD14+ monocytes—eliminating the need for complex flow cytometry equipment and delivering research-grade cell populations for all downstream applications.

Core Product Advantages

Key Research Applications

1. Monocyte/Macrophage Functional Research: Isolate CD14+ monocytes for in vitro differentiation into M1/M2 macrophages or dendritic cells; study phagocytosis, antigen presentation and inflammatory cytokine secretion.
2. Immunology and Disease Model Construction: Build in vitro immune inflammation models; study CD14+ cell roles in infections, autoimmune diseases, atherosclerosis and tumor microenvironment remodeling.
3. Cell Therapy and Regenerative Medicine: Use CD14+ monocytes as precursor cells for induced pluripotent stem cell (iPSC) generation, tissue engineering and immunomodulatory cell therapy research.
4. Clinical Biomarker Discovery: Isolate CD14+ cells from patient clinical samples to study phenotypic/functional changes in monocytes under disease states; identify novel immune biomarkers.
5. Omics and Single-Cell Research: Provide high-purity CD14+ cell populations for single-cell RNA sequencing, transcriptomics, proteomics and epigenomics analysis.

2. PKCζ in ADPKD: Molecular Mechanisms and Recombinant Rabbit Monoclonal Antibody Applications

2.1 ADPKD Pathogenesis and the Role of PKCζ

Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the most common inherited kidney disorder, caused by mutations in PKD1 (85% of cases) or PKD2 (15% of cases), leading to progressive bilateral renal cyst formation, parenchymal loss and end-stage renal disease. PKD1 encodes polycystin-1, a transmembrane glycoprotein with a cytoplasmic C-terminus that acts as a key regulatory hub for renal epithelial cell polarity, ciliogenesis and calcium signaling.

PKCζ—an atypical, PI3K/PDK1-dependent PKC isoform—plays a non-redundant role in ADPKD pathogenesis: it directly binds to the cytoplasmic C-terminus of polycystin-1 and mediates site-specific phosphorylation, a post-translational modification essential for maintaining polycystin-1 structural and functional integrity. Notably, PKCζ expression is significantly reduced in ADPKD patient tissues and preclinical animal models, and its dysfunction disrupts polycystin-1 signaling, leading to impaired epithelial polarity, abnormal cilia function and renal cyst initiation/progression.

2.2 PKCζ-Mediated Polycystin-1 Phosphorylation: A Key Regulator of Renal Epithelial Homeostasis

PKCζ modulates ADPKD through isoform-specific regulation of polycystin-1 and downstream signaling pathways:

1. PKCζ specifically interacts with polycystin-1’s cytoplasmic C-terminus and phosphorylates conserved serine/threonine residues, stabilizing polycystin-1 and enabling its interaction with other signaling molecules (e.g., polycystin-2).
2. PKCζ is a master regulator of epithelial cell polarity—one of the most dysregulated processes in ADPKD—controlling apical-basal membrane organization and tight junction formation in renal tubular cells.
3. PKCζ regulates ciliogenesis in renal epithelial cells; defective primary cilia (a hallmark of ADPKD) are directly linked to impaired PKCζ-polycystin-1 signaling.
4. PKCζ interacts with key metabolic and inflammatory signaling molecules (NF-κB, AMPK) in the kidney, amplifying its regulatory role in renal cell survival, metabolism and fibrosis—all pathological features of ADPKD.

2.3 PKCζ Recombinant Rabbit Monoclonal Antibody (S0B0802): Core Advantages and Research Applications

ANT BIO PTE. LTD.’s PKCζ Recombinant Rabbit mAb (S-1306-34, S0B0802) is a highly specific, high-affinity antibody engineered on the S-RMab® recombinant rabbit monoclonal platform—designed for isoform-specific analysis of PKCζ function in ADPKD and all PKCζ-mediated signaling pathways. It exhibits no cross-reactivity with other PKC isoforms (classical: α/β/γ; novel: δ/ε/η) and is reactive with human, mouse and rat PKCζ, enabling cross-species preclinical and translational research.

Core Product Advantages

1. Isoform-Specific Recognition: Targets PKCζ’s unique structural domains; minimal cross-reactivity with all other PKC family members, ensuring accurate analysis of PKCζ-specific signaling.
2. Pan-Species Reactivity: Recognizes PKCζ in human, mouse and rat; enables direct translation of mechanistic findings from preclinical animal models to human patient tissue studies.
3. Multi-Platform Utility: Optimized for Western Blot (WB), Immunoprecipitation (IP), Immunofluorescence (IF) and Immunohistochemistry (IHC); detects a ~70–80 kDa band in WB and precise subcellular localization (membrane/nuclear/cytoplasmic) in IF/IHC.
4. Superior Affinity and Stability: High-affinity recombinant rabbit monoclonal antibody enables sensitive detection of low-abundance PKCζ; rigorous quality control ensures batch-to-batch consistency and long-term stability for long-term research projects.
5. Phosphorylation and Interaction Analysis: Ideal for IP-based protein interaction studies (e.g., PKCζ-polycystin-1 complex capture) and paired with phospho-specific PKCζ antibodies to analyze kinase activity dynamics.

Key Research Applications in ADPKD and Beyond

1. ADPKD Mechanistic Research: Quantify PKCζ expression in ADPKD renal tissue; map PKCζ subcellular localization in cyst-forming renal epithelial cells; validate the PKCζ-polycystin-1 interaction and phosphorylation.
2. Therapeutic Response Monitoring: Evaluate the efficacy of PKCζ activators (e.g., fingolimod) in restoring PKCζ expression/activity in ADPKD preclinical models.
3. Atypical PKC Signaling: Study PDK1-mediated PKCζ activation and downstream signaling (e.g., NF-κB) in cell polarity, proliferation and survival.
4. Immune/Inflammatory Signaling: Dissect PKCζ’s role in TCR/BCR/TLR downstream pathways; analyze cross-talk between PKCζ and CD14/TLR4 innate immune signaling.
5. Metabolism and Cancer Research: Study PKCζ’s role in insulin signaling/GLUT4 translocation (diabetes) and context-dependent pro/anti-tumor functions in cancer cell proliferation, invasion and metastasis.

2.4 PKCζ Activation as a Novel Therapeutic Strategy for ADPKD

Restoring PKCζ function via pharmacological activation is a promising repurposed therapeutic strategy for ADPKD:

• Fingolimod (an FDA-approved immunomodulator) regulates ceramide levels—a known PKCζ activator—and exerts robust cyst-reducing and anti-fibrotic effects in ADPKD animal models dependent on PKCζ expression.
• This discovery validates PKCζ as a novel, actionable therapeutic target for ADPKD, offers a fast-track repurposing strategy for an existing drug, and defines the ceramide-PKCζ-polycystin-1 axis as a new molecular pathway for ADPKD therapy.
• The PKCζ recombinant rabbit monoclonal antibody (S0B0802) is the gold-standard tool for validating fingolimod’s on-target activity and for screening novel, renal-specific PKCζ activators with improved therapeutic potential.

Brand Mission of ANT BIO PTE. LTD.

At ANT BIO PTE. LTD., our core mission is to empower breakthroughs in immunology, nephrology, cell biology and translational drug development by delivering innovative, high-performance life science reagents and comprehensive solutions. As a leading global provider of research tools, we have built three specialized, complementary sub-brands that cover the full spectrum of life science research needs, creating a seamless one-stop procurement experience for academic researchers, biotech companies, pharmaceutical institutions and translational research labs worldwide:

• Starter: Our flagship sub-brand for antibodies and affinity purification/cell isolation tools, offering isoform-specific recombinant rabbit monoclonal antibodies (e.g., PKCζ S0B0802), MagSep™ cell sorting kits (e.g., CD14+ S0D3008), NA/LE grade functional antibodies and cell isolation reagents. Starter is dedicated to engineering highly specific, rigorously validated tools that enable the dissection of complex cell signaling pathways and the isolation of high-purity, functional cell populations for all areas of cell and disease biology.
• Absin: Our core sub-brand for general life science reagents and kits, providing cutting-edge OneStep ELISA Kits (cytokine, immunoglobulin), IHC detection kits, sample preparation reagents and cell culture media. Absin engineers user-friendly, pre-optimized assay kits that streamline quantitative immunoassay workflows and deliver reliable, accurate results for high-throughput research.
• UA: Specializes in high-purity, high-activity recombinant proteins and expression vectors, including recombinant cytokines, antibody heavy/light chain expression constructs, immunomodulatory proteins and gene editing vectors. UA enables seamless experimental design for protein expression, antibody engineering, cell therapy research and recombinant protein production.

We are committed to addressing the most pressing technical challenges in modern life science research—from isolating high-purity immune cells to dissecting isoform-specific molecular signaling pathways in human disease. By combining innovative tool design, rigorous validation protocols, standardized production and customer-centric scientific support, we translate technological innovation into research efficiency and breakthroughs for the global life science community. Our ultimate goal is to be the trusted global partner of researchers and biopharmaceutical professionals worldwide, empowering them to push the boundaries of disease research and drive unprecedented progress in medical science and human health.

Related Product List: CD14+ Cell Sorting Kits & PKCζ Antibody

All ANT BIO PTE. LTD. Starter products are rigorously validated for technical performance, specificity and batch-to-batch consistency. Each product is accompanied by a comprehensive technical documentation package (experimental protocols, validation data, troubleshooting guides) and our expert technical team provides one-on-one support for experimental design, optimization and downstream application development.

CD14 Positive Cell Sorting Kits (Starter MagSep™ Series)

PKCζ Recombinant Rabbit Monoclonal Antibody

For detailed product specifications, bulk pricing, custom cell isolation kit development, phospho-specific PKC antibody generation, or free sample testing requests, please visit the official website of ANT BIO PTE. LTD. or contact our global sales team for a personalized quote and professional technical consultation. Our experienced technical team also provides customized support for CD14+ cell functional research, ADPKD preclinical model validation and PKC signaling pathway analysis.

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ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs

At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.