Anti-Human CD14 [Clone UCHM-1] — Purified in vivo GOLD™ Functional Grade

Clone UCHM-1 is primarily used in in vivo mouse experiments to target human CD14, most commonly in humanized or xenograft mouse models where it can detect, deplete, or functionally block human monocytes and macrophages.

Key in vivo applications of clone UCHM-1 in mice include:

• Depletion of human monocytes/macrophages: UCHM-1 is used to selectively reduce or eliminate human CD14+ cells from humanized or chimeric mice to assess the role of these cells in immune responses or disease models.
• Functional blocking of CD14: By binding the CD14 receptor, UCHM-1 can block its function, such as its role in lipopolysaccharide (LPS) recognition and inflammatory signaling, to study CD14-mediated pathways in vivo.
• Detection/quantification of human CD14+ cells: Flow cytometry using UCHM-1 allows the identification and enumeration of human monocytes/macrophages within mouse tissues, useful for evaluating human cell engraftment or trafficking.
• Immunohistochemistry and immunofluorescence: UCHM-1 can be used to visualize the tissue localization of human monocytes/macrophages in mouse organs, aiding studies of cell distribution and microenvironmental interactions.

Important notes:

• Species specificity: UCHM-1 is specific for human CD14 and does not cross-react with mouse CD14, which is why it is particularly suited for use in mice engrafted with human immune cells.
• Model systems: These applications are mostly relevant to humanized mouse models (mice engrafted with human peripheral blood mononuclear cells, stem cells, or tissues) or xenograft systems where the interaction between human immune cells and murine or human tumor tissues is studied.

Summary of common in vivo UCHM-1 applications:

• Depletion of human CD14+ monocytes/macrophages in mice
• Functional blockade of human CD14 signaling
• Flow cytometric quantification of human monocytes/macrophages in chimeric mice
• Immunohistological mapping of human CD14+ cells in mouse tissues

These applications allow researchers to dissect the role of human monocyte/macrophage lineages in immunity, inflammation, infection, and tumor biology within the controlled environment of mouse models.

In the literature, UCHM-1 (a monoclonal antibody targeting CD14) is often used in combination with other antibodies to analyze immune cells, particularly in flow cytometry and inflammation studies. Here are some commonly used antibodies or proteins that are used with UCHM-1:

Antibodies Used with UCHM-1

1. CD3 Antibody: This targets the CD3 complex, which is part of the T cell receptor complex. CD3 and CD14 are used together to differentiate between T cells and monocytes/macrophages.

2. CD11b Antibody: This recognizes the CD11b antigen, which is found on monocytes, macrophages, and granulocytes. CD11b is often used alongside CD14 to examine myeloid cell populations.

3. CD16 Antibody: This targets the CD16 antigen present on neutrophils and some monocytes. It is used to identify these cell types in conjunction with CD14.

4. TLR4 Antibody: This recognizes the Toll-like receptor 4, which is involved in the recognition of LPS from gram-negative bacteria. TLR4 is sometimes studied in combination with CD14 due to their roles in the immune response to LPS.

5. HLA-DR Antibody: This targets the major histocompatibility complex class II molecule, often used in combination with CD14 to study antigen-presenting cells like dendritic cells and macrophages.

Proteins and Receptors

1. Lipopolysaccharide (LPS): Although not an antibody, LPS is often used in studies involving CD14 due to CD14's role as a co-receptor for LPS recognition.

2. TLR4: Similar to LPS, TLR4 is a receptor that works with CD14 in the recognition of LPS. It is a key component in the innate immune response and is frequently studied alongside CD14.

These antibodies and proteins are used to understand the roles of myeloid cells and the mechanisms of immune responses, particularly against bacterial components like LPS.

Clone UCHM-1 is a monoclonal antibody directed against human CD14, a key molecule in innate immunity, and its citations in scientific literature have produced several important findings:

• UCHM-1 inhibits LPS-induced inflammatory responses in immune cells. Specifically, anti-CD14 monoclonal antibodies (mAbs) of the UCHM-1 clone attenuate LPS-induced priming of human polymorphonuclear neutrophils (PMNs), as seen by a reduction in fMLP-triggered reactive oxygen species (ROS) production.
• UCHM-1 identifies CD14 as essential for LPS-driven PMN priming and ROS release. The effect is specific to CD14 blockade, as antibodies against other targets (e.g., TLR4 or CD11b) do not produce the same reduction in ROS.
• UCHM-1 selectively targets a critical epitope on CD14 for endotoxin signaling. The UCHM-1 clone (and related antibody MY4) binds the SAVEVEIHAGG epitope, crucial for LPS-induced signal transduction, unlike other anti-CD14 clones such as 63D3 or biG6 that fail to block this pathway.
• Blockade of CD14 using UCHM-1 or its fragments has potential therapeutic effects in LPS-mediated inflammation. Antibody-mediated inhibition reduces ROS production, which could be beneficial in conditions like cardiovascular diseases linked to endotoxin-driven inflammation.
• UCHM-1 is valuable for studying and modulating inflammatory pathways and immune cell function. It is widely used in research to explore the function of CD14 and the mechanisms of innate immune recognition.

Additional technical details:

• UCHM-1 recognizes a specific cell surface glycoprotein (CD14, Mr 55 kDa) expressed on most peripheral blood monocytes and granulocytes.
• CD14 acts as a co-receptor for LPS and is involved in binding apoptotic cells and modulating immune processes.
• UCHM-1’s efficacy in flow cytometry is demonstrated by its strong and specific binding to CD14-expressing cells.

In summary, clone UCHM-1 citations consistently highlight its key role in defining CD14’s contribution to LPS-mediated cell activation, its specificity for a critical functional epitope, and its utility in both basic and translational research targeting inflammatory responses.

Dosing regimens for clone UCHM-1 (mouse anti-human CD14) are not universally established for in vivo use in mouse models, as this clone is most frequently cited for applications such as flow cytometry, immunohistochemistry, and ex vivo cell enumeration, rather than therapeutic or depleting antibody studies in mice. Most product and literature references recommend specific volumes or concentrations for cell staining, typically around 10 μL per 10^6 cells for flow cytometry.

Key details:

• Primary application: Flow cytometry and immunohistology for detection of human CD14+ cells within xenograft or humanized mouse models.
• Suggested staining dose: 10 μL (of working dilution) per 10^6 target cells in 100 μL for FACS analysis.
• No standard in vivo depletion or blocking regimen cited in the available research or manufacturer documentation.

Context and limitations:

• In contrast to checkpoint blockade antibodies used for functional studies in mice (such as anti-PD-1, anti-CTLA-4, etc.), for which detailed dosing regimens are well established (e.g., 100–250 μg per mouse every 3 days via intraperitoneal injection), UCHM-1 is not typically used for repeated systemic in vivo administration or functional depletion of CD14+ cells in mouse models.
• If considering UCHM-1 for in vivo blocking or depletion of human CD14+ cells in a humanized mouse model, regimen would need to be empirically determined, most likely starting in the range of 100–250 μg/mouse, modeled after protocols for other mouse anti-human antibodies, with close monitoring for efficacy and toxicity. No directly cited successful examples for in vivo dosing of UCHM-1 were found in the provided sources.
• As a mouse IgG2a, UCHM-1 may engage mouse Fc receptors, but efficacy for depleting human CD14+ cells in vivo must be validated for each model.

Summary table:

If your research requires in vivo depletion or functional blockade with UCHM-1, pilot studies and titration in your specific mouse model (e.g., humanized mice engrafted with human immune cells) would be necessary, given the absence of standardized dosing recommendations in the literature.