Anti-Human CD14 (61D3) ColorWheel^® Dye-Ready mAb

ColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel^® antibody with any ColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel^® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel^® technology requires both ColorWheel^® antibody and ColorWheel^® dye for Flow Cytometry application.

Monocyte differentiation antigen CD14 (UniProt: P08571; also known as Myeloid cell-specific leucine-rich glycoprotein, CD14) is encoded by the CD14 gene (Gene ID: 929) in human. CD14 is a GPI-anchor membrane glycoprotein that is synthesized with a signal peptide (aa 1-19) and a propeptide (aa 346-375) that are cleaved off to generate the mature form. The mature form contains 11 leucine-rich repeats (LRR) that are required for responses to smooth LPS. It serves as a co-receptor for several Toll-like receptors. Together with TLR4 and MD-2, it forms the multi-receptor complex that recognizes Lipopolysaccharide (LPS) on the cell membrane. In the presence of LPS, CD14 and TLR4-MD2 are brought into close proximity inside lipid rafts and after translocation into the lipid rafts, TLR4 in the plasma membrane activates the TIRAP-MyD88-dependent pathway that leads to the first wave of NF-kB activation. CD14 also acts as a co-receptor for TLR2:TLR6 heterodimer in response to diacylated lipopeptides and for TLR2:TLR1 heterodimer in response to triacylated lipopeptides. These clusters trigger signaling from the cell surface and subsequently are targeted to the Golgi in a lipid-raft dependent pathway. CD14 expression has been reported on cells of both hematopoietic and non-hematopoietic origin. A secreted form of CD14 has also reported that arises from the cleavage of the GPI anchor. CD14 is expressed on macrophages and a strong expression has been reported on the surface of monocytes. Its expression on monocytes is induced by 27-hydroxycholesterol that primes monocytes/macrophages to accelerate LPS-mediated inflammatory reaction. CD14-deficient macrophages display a significantly reduced sensitivity to low concentrations of LPS compared to wild-type cells and CD14 deficient mice do not develop septic shock after exposure to LPS or Gram-negative bacteria. (Ref.: Zanoni, I., and Granucci, F. (2013). Front. Cell. Infect. Microbiol. 3; 32; Wright, SD., et al. (1990). Science. 249(4975); 1431-1433).

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Human peripheral monocytes.

Quality Control Testing

Evaluated by Flow Cytometry in human peripheral blood monocytes (PBMC).

Flow Cytometry Analysis: Staining of one million human peripheral blood mononuclear cells (PBMC) was performed using 5 μL of a 1:1 mixture of Cat. No. CWA-1089, Anti-Human CD14 (61D3) ColorWheel^® Dye-Ready mAb and Cat. No. CWD-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent PE-conjugated Mouse IgG1 isotype control.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user

Clone 61D3 is a mouse monoclonal antibody that detects human Monocyte differentiation antigen CD14 (CD14).

Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.

1.0 mg/mL after reconstitution at 25 μL in PBS. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C. Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial. Reconstitute each vial by adding 25 μL PBS. Protect from light.

ColorWheel is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.