CD14 antibody

Catalog No. ABIN967379

Product Details anti-CD14 Antibody

Target: CD14

Reactivity: Mouse

Host: Rat

Clonality: Monoclonal

Conjugate: This CD14 antibody is un-conjugated

Application: Flow Cytometry (FACS), Western Blotting (WB)

Brand: BD Pharmingen™

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Sodium azide is a reversible inhibitor of oxidative metabolism, therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE™ (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Recombinant Mouse CD14

Clone: RmC5-3

Isotype: IgG1 kappa

Application Details

Application Notes: Caution: Our studies demonstrate that Mouse Fc Block™ anti-CD16/CD32 mAb 2.4G2 and antibodies of rat IgG2b isotype may interfere with the reactivity of rmC5-3 antibody in a concentration-dependent manner. For inhibition of non-specific reactivity, we recommend use of purified mouse IgG at 10-100-fold excess. Other reported applications include western blot analysis. For in vitro and in vivo blocking of LPS binding to CD14, we recommend the NA/LE™ format of the alternate anti-mouse CD14 mAb 4C1/CD14. Use of rmC5-3 antibody for immunohistochemical staining has been reported, however, we have been unable to reproduce those results.

Comment:

Related Products: ABIN967379

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 0.5 mg/mL

Buffer: Aqueous buffered solution containing ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: 4 °C

Storage Comment: Store undiluted at 4° C.

References for anti-CD14 antibody (ABIN967379)

Fearns, Loskutoff: "Role of tumor necrosis factor alpha in induction of murine CD14 gene expression by lipopolysaccharide." in: Infection and immunity, Vol. 65, Issue 11, pp. 4822-31, (1997) (PubMed).

Pulendran, Lingappa, Kennedy, Smith, Teepe, Rudensky, Maliszewski, Maraskovsky: "Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 159, Issue 5, pp. 2222-31, (1997) (PubMed).

Takakuwa, Knopf, Sing, Carsetti, Galanos, Freudenberg: "Induction of CD14 expression in Lpsn, Lpsd and tumor necrosis factor receptor-deficient mice." in: European journal of immunology, Vol. 26, Issue 11, pp. 2686-92, (1997) (PubMed).

Ziegler-Heitbrock: "Heterogeneity of human blood monocytes: the CD14+ CD16+ subpopulation." in: Immunology today, Vol. 17, Issue 9, pp. 424-8, (1997) (PubMed).

Mahnke, Becher, Ricciardi-Castagnoli, Luger, Schwarz, Grabbe: "CD14 is expressed by subsets of murine dendritic cells and upregulated by lipopolysaccharide." in: Advances in experimental medicine and biology, Vol. 417, pp. 145-59, (1997) (PubMed).

Haziot, Ferrero, Köntgen, Hijiya, Yamamoto, Silver, Stewart, Goyert: "Resistance to endotoxin shock and reduced dissemination of gram-negative bacteria in CD14-deficient mice." in: Immunity, Vol. 4, Issue 4, pp. 407-14, (1996) (PubMed).

Takamatsu, Nakashima, Nakano: "Modulation of endotoxin-induced histamine synthesis by cytokines in mouse bone marrow-derived macrophages." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 156, Issue 2, pp. 778-85, (1996) (PubMed).

Takezawa, Watanabe, Akaike: "Direct evidence of macrophage differentiation from bone marrow cells in the liver: a possible origin of Kupffer cells." in: Journal of biochemistry, Vol. 118, Issue 6, pp. 1175-83, (1996) (PubMed).

Fearns, Kravchenko, Ulevitch, Loskutoff: "Murine CD14 gene expression in vivo: extramyeloid synthesis and regulation by lipopolysaccharide." in: The Journal of experimental medicine, Vol. 181, Issue 3, pp. 857-66, (1995) (PubMed).

Matsuura, Ishida, Setoguchi, Higuchi, Akizuki, Yamamoto: "Upregulation of mouse CD14 expression in Kupffer cells by lipopolysaccharide." in: The Journal of experimental medicine, Vol. 179, Issue 5, pp. 1671-6, (1994) (PubMed).

Nasu, Yoshida, Akizuki, Higuchi, Setoguchi, Yamamoto: "Molecular and physiological properties of murine CD14." in: International immunology, Vol. 3, Issue 2, pp. 205-13, (1991) (PubMed).

Ferrero, Hsieh, Francke, Goyert: "CD14 is a member of the family of leucine-rich proteins and is encoded by a gene syntenic with multiple receptor genes." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 145, Issue 1, pp. 331-6, (1990) (PubMed).

Target Details for CD14

Target: CD14

Alternative Name: CD14 (CD14 Products)

Synonyms: CD14 molecule antibody, CD14 antigen antibody, CD14 antibody, Cd14 antibody

Background: The rmC5-3 antibody reacts with residues 308-322 of the hydrophilic region of mouse CD14. CD14 is a 53-55 kDa glycosyl phosphatidyl inositol (GPI)-linked glycoprotein belonging to the leucine-rich glycoprotein repeat superfamily of cell-surface proteins. It is a receptor for the complex of lipopolysaccharide (LPS or endotoxin, from gram-negative bacteria) with LPS-binding protein (LBP, a plasma protein). It is involved in the development of endotoxic shock and LPS-stimulated bone resorption, and promotes, possibly indirectly, bacterial dissemination. Flow cytometric analysis demonstrates that rmC5-3 antibody stains J774A.1 (mouse macrophage line), WEHI-265.1 (mouse monocytic line), peritoneal resident macrophages, Kupffer cells, and cultured bone marrow-derived macrophages and dendritic cells, but not unstimulated splenic macrophages, dendritic cells, neutrophils, or blood monocytes. This staining pattern is similar to that of the alternate anti-mouse CD14 mAb 4C1/CD14, which recognizes a different CD14 epitope, and differs from that of the human, where CD14 expression is characteristic of circulating monocytes and neutrophils. Therefore, data suggests that CD14 expression by leukocyte populations may differ in mice and humans. Peritoneal cells from naive mice, 3-day thioglycollate-elicited peritoneal exudate, as well as 4-hour LPS-activated peritoneal cells, contain a population of Mac-1 (CD11b)-high cells which double-stain with rmC5-3 antibody. Levels of CD14 expression on Kupffer cells and bone marrow-derived macrophages and dendritic cells of LPS-sensitive mice are increased by in vivo and in vitro LPS treatments, an effect which may be mediated by TNF-alpha. Preliminary evidence suggests that CD14 may be up-regulated on mouse blood neutrophils. In agreement with the observations that CD14 is shed from activated human and mouse monocytes, rmC5-3 mAb detects soluble CD14 in the serum of LPS-treated mice in a time-dependent manner. This antibody is routinely tested by flow cytometric analysis.

Pathways: TLR Signaling, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Toll-Like Receptors Cascades