Human CD14 Antibody Summary
Accession # P08571
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of CD14 in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) were stained with Mouse Anti-Human CD14 Monoclonal Antibody (Catalog # MAB3832R) followed by Goat anti-Mouse IgG APC-conjugated Secondary Antibody (Catalog # F0101B). View our protocol for Staining Membrane-associated Proteins.
TNF-alpha Secretion Induced by LPS and Neutralization by Human CD14 Antibody. Lipopolysaccharide (LPS) stimulates TNF-alpha secretion in the THP-1 human acute monocytic leukemia cell line in a dose-dependent manner (orange line). TNF-alpha secretion elicited by LPS is neutralized (green line) by increasing concentrations of Mouse Anti-Human CD14 Monoclonal Antibody (Catalog # MAB3832R). At 10 μg/mL, this anti-human CD14 antibody will neutralize 60% of 0.5 ng/mL LPS-induced TNF-alpha secretion.
Preparation and Storage
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
CD14 is a 55 kDa cell surface glycoprotein that is preferentially expressed on monocytes/macrophages. The human CD14 cDNA encodes a 375 amino acid (aa) residue precursor protein with a 19 aa signal peptide and a C-terminal hydrophobic region characteristic for glycosylphosphatidyinositol (GPI)-anchored proteins. Human CD14 has four potential N-linked glycosylation sites and also bears O-linked carbohydrates. The amino acid sequence of human CD14 is approximately 65% identical with the mouse, rat, rabbit, and bovine proteins. CD14 is a pattern recognition receptor that binds lipopolysaccharides (LPS) and a variety of ligands derived from different microbial sources. The binding of CD14 with LPS is catalyzed by LPS-binding protein (LBP). The toll-like-receptors have also been implicated in the transduction of CD14-LPS signals. Similar to other GPI-anchored proteins, soluble CD14 can be released from the cell surface by phosphatidyinositol-specific phospholipase C. Soluble CD14 has been detected in serum and body fluids. High concentrations of soluble CD14 have been shown to inhibit LPS-mediated responses. However, soluble CD14 can also potentiate LPS response in cells that do not express cell surface CD14.
- Wright, S.D. et al. (1990) Science 249:1431.
- Pugin, J. et al. (1993) Proc. Natl. Acad. Sci. USA 90:2744.
- Beutler, B. (2000) Current Opinion in Immunology 12:20.
- Stelter, F. (2000) Chem. Immunol. 74:25.
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