categories: CD and Related Antigens (Human), CD Antigens & MHC (Veterinary)

Mouse Monoclonal to CD14 / LPS-receptor

MEM-18 (IgG1)

Product No.	Form	Quantity	Price
11-212-C025	purified	0.025 mg	choose region
11-212-C100	purified	0.1 mg	choose region
12-212-C100	low endotoxin	0.1 mg	choose region
F2-212-C100	F(ab')2 fragment	0.1 mg	choose region
1B-212-C025	biotin	0.025 mg	choose region
1B-212-C100	biotin	0.1 mg	choose region
PB-212-T025	Pacific Blue™	25 tests	choose region
PB-212-T100	Pacific Blue™	100 tests	choose region
1F-212-T025	FITC	25 tests	choose region
1F-212-T100	FITC	100 tests	choose region
1P-212-T025	PE	25 tests	choose region
1P-212-T100	PE	100 tests	choose region
1A-212-T025	APC	25 tests	choose region
1A-212-T100	APC	100 tests	choose region
A7-212-T025	Alexa Fluor® 700	25 tests	choose region
A7-212-T100	Alexa Fluor® 700	100 tests	choose region

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Background

CD14 is a 55 kDa GPI-anchored glycoprotein, constitutively expressed on the surface of mature monocytes, macrophages, and neutrophils, where serves as a multifunctional lipopolysaccharide receptor; it is also released to the serum both as a secreted and enzymatically cleaved GPI-anchored form. CD14 binds lipopolysaccharide molecule in a reaction catalyzed by lipopolysaccharide-binding protein (LBP), an acute phase serum protein. The soluble sCD14 is able to discriminate slight structural differences between lipopolysaccharides and is important for neutralization of serum allochthonous lipopolysaccharides by reconstituted lipoprotein particles. CD14 affects allergic, inflammatory and infectious processes.

Entrez Gene (human): 5q31.1

Specificity

The antibody MEM-18 reacts with CD14, a 53-55 kDa GPI (glycosylphosphatidylinositol)-linked membrane glycoprotein expressed on monocytes, macrophages and weakly on granulocytes; also expressed by most tissue macrophages. In human, the epitope recognized by MEM-18 is located between amino acids 57-64.
HLDA III; WS Code M 253
HLDA IV; WS Code M 314
HLDA V; WS Code M MA087
HLDA VI; WS Code M MA95

Immunogen

A crude mixture of human urinary proteins precipitated by ammonium sulphate from the urine of a patient suffering from proteinuria.

Species Reactivity: Human Non-Human Primates Negative Species:	Applications: Flow Cytometry Recommended dilution:4 μg/ml Immunoprecipitation Western Blotting Application note: Non-reducing conditions. ELISA The antibody MEM-18 has been tested as the detection antibody in a sandwich ELISA for analysis of human CD14 in combination with antibody B-A8 (cat. no. 11-304-C100). Functional Application The antibody MEM-18 completely blocks binding of fluorescein (FITC) – labeled bacterial LPS to the monocyte surface and it also blocks the binding of CD14 to the extracellular TLR2 domain. Usage note: Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.	General references Juan TS, Hailman E, Kelley MJ, Wright SD, Lichenstein HS: Identification of a domain in soluble CD14 essential for lipopolysaccharide (LPS) signaling but not LPS binding. J Biol Chem. 1995 Jul 21;270(29):17237-42. [Abstract] [Full Text] Lodrup Carlsen KC, Granum B: Soluble CD14: role in atopic disease and recurrent infections, including otitis media. Curr Allergy Asthma Rep. 2007 Nov;7(6):436-43. [Abstract] Asai Y, Makimura Y, Kawabata A, Ogawa T: Soluble CD14 Discriminates Slight Structural Differences between Lipid As That Lead to Distinct Host Cell Activation. J Immunol. 2007 Dec 1;179(11):7674-83. [Abstract] Fernández-Real JM, Broch M, Richart C, Vendrell J, López-Bermejo A, Ricart W: CD14 monocyte receptor, involved in the inflammatory cascade, and insulin sensitivity. J Clin Endocrinol Metab. 2003 Apr;88(4):1780-4. [Abstract] [Full Text] Product Specific References Bazil V, Horejsi V, Baudys M, Kristofova H, Strominger JL, Kostka W, Hilgert I: Biochemical characterization of a soluble form of the 53-kDa monocyte surface antigen. Eur J Immunol. 1986 Dec;16(12):1583-9. [Abstract] Leukocyte Typing III., McMichael A.J. et al. (Eds.), Oxford University Press (1987). Bazil V, Baudys M, Hilgert I, Stefanova I, Low MG, Zbrozek J, Horejsi V: Structural relationship between the soluble and membrane-bound forms of human monocyte surface glycoprotein CD14. Mol Immunol. 1989 Jul;26(7):657-62. [Abstract] Leukocyte Typing IV., Knapp W. et al. (Eds.), Oxford University Press (1989). Leukocyte Typing V., Schlossman S. et al. (Eds.), Oxford University Press (1995). Leukocyte Typing VI., Kishimoto T. et al. (Eds.), Garland Publishing Inc. (1997). Iwaki D, Nishitani C, Mitsuzawa H, Hyakushima N, Sano H, Kuroki Y: The CD14 region spanning amino acids 57-64 is critical for interaction with the extracellular Toll-like receptor 2 domain. Biochem Biophys Res Commun. 2005 Mar 4;328(1):173-6. [Abstract] Angel CE, Lala A, Chen CJ, Edgar SG, Ostrovsky LL, Dunbar PR: CD14+ antigen-presenting cells in human dermis are less mature than their CD1a+ counterparts. Int Immunol. 2007 Nov;19(11):1271-9. [Abstract] [Full Text] Drbal K, Moertelmaier M, Holzhauser C, Muhammad A, Fuertbauer E, Howorka S, Hinterberger M, Stockinger H, Schütz GJ: Single-molecule microscopy reveals heterogeneous dynamics of lipid raft components upon TCR engagement. Int Immunol. 2007 May;19(5):675-84. [Abstract] [Full Text] Weiss TS, Lichtenauer M, Kirchner S, Stock P, Aurich H, Christ B, Brockhoff G, Kunz-Schughart LA, Jauch KW, Schlitt HJ, Thasler WE. Hepatic progenitor cells from adult human livers for cell transplantation. Gut. 2008 Aug;57(8):1129-38. [Abstract] [Full Text] Stöckl J, Majdic O, Fischer G, Maurer D, Knapp W: Monomorphic molecules function as additional recognition structures on haptenated target cells for HLA-A1-restricted, hapten-specific CTL. J Immunol. 2001 Sep 1;167(5):2724-33. [Abstract] [Full Text] Funda DP, Tucková L, Farré MA, Iwase T, Moro I, Tlaskalová-Hogenová H: CD14 is expressed and released as soluble CD14 by human intestinal epithelial cells in vitro: lipopolysaccharide activation of epithelial cells revisited. Infect Immun. 2001 Jun;69(6):3772-81. [Abstract] [Full Text] Sing A, Rost D, Tvardovskaia N, Roggenkamp A, Wiedemann A, Kirschning CJ, Aepfelbacher M, Heesemann J: Yersinia V-antigen exploits toll-like receptor 2 and CD14 for interleukin 10-mediated immunosuppression. J Exp Med. 2002 Oct 21;196(8):1017-24. [Abstract] [Full Text] And other.

Species Reactivity:

Human
Non-Human Primates

Negative Species:

Applications:

Flow Cytometry
Recommended dilution:4 μg/ml
Immunoprecipitation
Western Blotting
Application note: Non-reducing conditions.
ELISA
The antibody MEM-18 has been tested as the detection antibody in a sandwich ELISA for analysis of human CD14 in combination with antibody B-A8 (cat. no. 11-304-C100).
Functional Application
The antibody MEM-18 completely blocks binding of fluorescein (FITC) – labeled bacterial LPS to the monocyte surface and it also blocks the binding of CD14 to the extracellular TLR2 domain.

Usage note:
Indicated dilutions are recommended starting points for use of this product. Working concentrations should be determined by the investigator.

General references

*Juan TS, Hailman E, Kelley MJ, Wright SD, Lichenstein HS: Identification of a domain in soluble CD14 essential for lipopolysaccharide (LPS) signaling but not LPS binding. J Biol Chem. 1995 Jul 21;270(29):17237-42. [Abstract] [Full Text]

*Lodrup Carlsen KC, Granum B: Soluble CD14: role in atopic disease and recurrent infections, including otitis media. Curr Allergy Asthma Rep. 2007 Nov;7(6):436-43. [Abstract]

*Asai Y, Makimura Y, Kawabata A, Ogawa T: Soluble CD14 Discriminates Slight Structural Differences between Lipid As That Lead to Distinct Host Cell Activation. J Immunol. 2007 Dec 1;179(11):7674-83. [Abstract]

*Fernández-Real JM, Broch M, Richart C, Vendrell J, López-Bermejo A, Ricart W: CD14 monocyte receptor, involved in the inflammatory cascade, and insulin sensitivity. J Clin Endocrinol Metab. 2003 Apr;88(4):1780-4. [Abstract] [Full Text]

Product Specific References

*Bazil V, Horejsi V, Baudys M, Kristofova H, Strominger JL, Kostka W, Hilgert I: Biochemical characterization of a soluble form of the 53-kDa monocyte surface antigen. Eur J Immunol. 1986 Dec;16(12):1583-9. [Abstract]

*Leukocyte Typing III., McMichael A.J. et al. (Eds.), Oxford University Press (1987).

*Bazil V, Baudys M, Hilgert I, Stefanova I, Low MG, Zbrozek J, Horejsi V: Structural relationship between the soluble and membrane-bound forms of human monocyte surface glycoprotein CD14. Mol Immunol. 1989 Jul;26(7):657-62. [Abstract]

*Leukocyte Typing IV., Knapp W. et al. (Eds.), Oxford University Press (1989).

*Leukocyte Typing V., Schlossman S. et al. (Eds.), Oxford University Press (1995).

*Leukocyte Typing VI., Kishimoto T. et al. (Eds.), Garland Publishing Inc. (1997).

*Iwaki D, Nishitani C, Mitsuzawa H, Hyakushima N, Sano H, Kuroki Y: The CD14 region spanning amino acids 57-64 is critical for interaction with the extracellular Toll-like receptor 2 domain. Biochem Biophys Res Commun. 2005 Mar 4;328(1):173-6. [Abstract]

*Angel CE, Lala A, Chen CJ, Edgar SG, Ostrovsky LL, Dunbar PR: CD14+ antigen-presenting cells in human dermis are less mature than their CD1a+ counterparts. Int Immunol. 2007 Nov;19(11):1271-9. [Abstract] [Full Text]

*Drbal K, Moertelmaier M, Holzhauser C, Muhammad A, Fuertbauer E, Howorka S, Hinterberger M, Stockinger H, Schütz GJ: Single-molecule microscopy reveals heterogeneous dynamics of lipid raft components upon TCR engagement. Int Immunol. 2007 May;19(5):675-84. [Abstract] [Full Text]

*Weiss TS, Lichtenauer M, Kirchner S, Stock P, Aurich H, Christ B, Brockhoff G, Kunz-Schughart LA, Jauch KW, Schlitt HJ, Thasler WE. Hepatic progenitor cells from adult human livers for cell transplantation. Gut. 2008 Aug;57(8):1129-38. [Abstract] [Full Text]

*Stöckl J, Majdic O, Fischer G, Maurer D, Knapp W: Monomorphic molecules function as additional recognition structures on haptenated target cells for HLA-A1-restricted, hapten-specific CTL. J Immunol. 2001 Sep 1;167(5):2724-33. [Abstract] [Full Text]

*Funda DP, Tucková L, Farré MA, Iwase T, Moro I, Tlaskalová-Hogenová H: CD14 is expressed and released as soluble CD14 by human intestinal epithelial cells in vitro: lipopolysaccharide activation of epithelial cells revisited. Infect Immun. 2001 Jun;69(6):3772-81. [Abstract] [Full Text]

*Sing A, Rost D, Tvardovskaia N, Roggenkamp A, Wiedemann A, Kirschning CJ, Aepfelbacher M, Heesemann J: Yersinia V-antigen exploits toll-like receptor 2 and CD14 for interleukin 10-mediated immunosuppression. J Exp Med. 2002 Oct 21;196(8):1017-24. [Abstract] [Full Text]

*And other.

For research use only. Not for drug, diagnostic or other use.

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Mouse IgG1 Isotype Control

Example Data

Fig. 1. Western Blotting

Fig. 1. Western Blotting analysis (non-reducing conditions) of over-expressed human CD14 using anti-CD14 (MEM-18).
Lane 1: whole cell lysate HEK 293 transfected with empty vector
Lane 2: tissue culture supernatant collected after cultivation of HEK 293 transfected with human CD14 cDNA
Lane 3: whole cell lysate of HEK 293 transfected with human CD14 cDNA

Fig. 2.

Fig. 2. Surface staining of human peripheral blood leukocytes using anti-human CD14 (clone MEM-18) FITC.

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