Anti-Human CD14 – Purified in vivo GOLD™ Functional Grade

Clone UCHM-1 is a mouse monoclonal antibody specifically directed against human CD14, and in in vivo mouse studies, it is primarily used to detect, deplete, or functionally block human monocytes and macrophages in humanized or xenograft mouse models.

In essential context:

• UCHM-1 does not bind murine CD14; it is specific to human (and cynomolgus) CD14. Therefore, it is useful only in in vivo mouse experiments where human immune cells are present, such as humanized mice engrafted with human hematopoietic stem cells or tissues.
• Applications include:
  • Flow cytometry of mouse tissues to identify and quantify human monocytes/macrophages in chimeric mice.
  • Immunohistochemistry/immunocytochemistry on tissue sections to localize human CD14+ cells within mouse organs.
  • In vivo cell depletion: When administered to humanized mice, UCHM-1 can be used for antibody-mediated depletion of human monocytes/macrophages, allowing functional studies of their role.
  • Functional blocking: UCHM-1 can inhibit CD14 function in vivo, enabling the investigation of CD14-mediated signaling and responses, such as LPS-induced immune activation.
• Mechanistic notes: In humanized mice, the presence and function of human monocytes/macrophages can be tracked and manipulated using UCHM-1, which is crucial for studying human immune responses, especially in infection, inflammation, or tumor xenograft models.

Key limitations:

• UCHM-1 does not recognize mouse CD14, so it is not useful in wild-type mice without human cell engraftment.
• It is most often used in research involving humanized mouse models, where accurate tracking or control of human immune cell populations is necessary.

Summary table for UCHM-1 use in mouse studies:

In summary: In mouse studies, UCHM-1 is a vital tool for interrogating the behavior and function of human monocytes/macrophages in vivo, but it is only applicable in models containing human (or cynomolgus) immune cells.

The correct storage temperature for sterile packaged clone UCHM-1 (a monoclonal anti-human CD14 antibody) is -20°C for long-term storage or +4°C (refrigerated) for short-term use. It should be kept undiluted and stored in a non–frost-free freezer to avoid freeze-thaw cycles if at -20°C. For periods shorter than a month, storage at +4°C is acceptable; for longer periods, -20°C is strongly recommended.

For conjugated forms of the antibody (e.g., PE-conjugated), some manufacturers recommend storage at 2–8°C (refrigerated) and protection from light. Always avoid room temperature storage for extended periods and repeated freeze-thawing, as this can reduce antibody efficacy.

• -20°C: For long-term storage (months to years).
• +4°C: For short-term storage (up to 1 month).
• Frost-free freezers should be avoided due to temperature fluctuations.
• Aliquoting is recommended to prevent loss of activity from repeated freeze-thaw cycles.

Always follow storage instructions on the specific antibody datasheet and consult the manufacturer if uncertain.

Commonly used antibodies or proteins that are paired with UCHM-1 (an anti-human CD14 antibody) in the literature primarily include markers that identify different leukocyte populations or related immune molecules. The most frequently reported combinations, especially in flow cytometry and immunophenotyping, are:

• CD3: A T cell marker, often conjugated to different fluorochromes, and commonly used alongside UCHM-1 to distinguish monocytes (CD14?) from T lymphocytes (CD3?).
• CD11b/CD18 (Mac-1): A marker for myeloid cells, such as monocytes and granulocytes, and can be used to further define myeloid subsets or to compare expression with CD14.
• TLR4: Toll-like receptor 4 is functionally linked with CD14 in innate immune signaling and is sometimes analyzed in combination with UCHM-1, especially in studies examining LPS responses.
• HLA-DR: A major histocompatibility complex class II molecule, often used with CD14 to assess monocyte activation and antigen-presenting capacity (not explicit in the search results but widely used in standard immunophenotyping panels).
• CD16: Sometimes used to distinguish monocyte subsets (e.g., classical CD14++CD16? vs. nonclassical CD14+CD16++ monocytes).

In addition, UCHM-1 is frequently used with various panel markers in:

• Flow cytometry: For leukocyte subset analysis (as demonstrated by its pairing with anti-CD3 in multiple references).
• Immunohistochemistry and immunocytochemistry: Sometimes with T cell, B cell, or other myeloid markers to characterize tissue or cellular infiltrates.

Summary table: Commonly co-used markers with UCHM-1 (anti-CD14)

Conclusion: UCHM-1 is frequently used in immune phenotyping panels together with antibodies against CD3, CD11b, CD16, and TLR4, among others, to define and analyze monocyte populations in human samples, especially in flow cytometry and inflammation research.

Based on the scientific literature, clone UCHM-1 is a monoclonal antibody that has been extensively studied for its interactions with CD14, revealing several key findings across different research areas.

Antibody Characteristics and Specificity

Clone UCHM-1 is a mouse anti-human CD14 monoclonal antibody with an IgG2a isotype that recognizes a cell surface antigen of approximately 55 kDa known as CD14. The antibody demonstrates specific binding patterns, showing strong expression on monocytes and macrophages in flow cytometry, weaker expression on granulocytes, and absence from stem cells and early differentiation state myeloid cells. In immunohistological studies, UCHM-1 positively stains monocytic cells, interfollicular tissue macrophages, dendritic reticulum cells, Kupffer cells, and sinus lining cells in the liver, while notably showing negative reactions with skin Langerhans cells.

Functional Effects on Cellular Responses

One of the most significant findings is that antibodies to the CD14 molecule, including UCHM-1, are known to induce oxidative burst formation. This biological activity has important implications for understanding immune cell activation and inflammatory responses. Additionally, research has demonstrated that anti-CD14 monoclonal antibodies using clone UCHM-1 can attenuate LPS-induced priming of polymorphonuclear neutrophils (PMNs), as evidenced by a decrease in fMLP-triggered reactive oxygen species (ROS) production.

Therapeutic and Research Applications

The UCHM-1 clone has proven valuable in studying inflammatory pathways and immune cell function. Its ability to modulate LPS-induced cellular responses makes it particularly useful for investigating endotoxin-mediated inflammation and the role of CD14 in innate immune recognition. The antibody's specific tissue distribution pattern also makes it a useful tool for identifying and characterizing different populations of immune cells in various tissues and experimental conditions.

Comparative Effectiveness Studies

Research comparing different anti-CD14 antibody clones has shown that the effectiveness of anti-CD14 UCHM-1 FcmIgG2a antibody against LPS-induced effects in human monocytes may be less evident compared to other clones, such as anti-CD14 MEM variants. This finding highlights the importance of epitope specificity and antibody engineering in determining therapeutic efficacy.

These findings collectively demonstrate that clone UCHM-1 serves as both a valuable research tool for studying CD14-mediated immune responses and a potential therapeutic agent for modulating inflammatory processes, though its effectiveness may vary depending on the specific experimental context and target cell populations.