Mouse anti-Human CD14 Antibody | anti-CD14 antibody

Mouse anti CD14

Cat. Num. AAA570362

• Reactivity: Human
• Applications: Immunofluorescence, Immunofluorescence, Flow Cytometry, Functional Assay
• Synonyms: CD14; Mouse anti CD14; anti-CD14 antibody

• Host: Mouse
• Reactivity: Human
• Isotype: IgG1
• Clone Number: MEM18
• Specificity: The CD14 mAb (clone MEM18) recognizes surface CD14 on human monocytes and macrophages as well as on neutrophils.; ; The sensitivity of MEM18 mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAb; dilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI; (mean fluorescence intensity). For this purpose, a mAb-concentration range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50 ul of leukocytes containing 10^7 cells/ml are stained with 20 ul mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection threshold. The final concentration of the product is then adjusted to be at least 3-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.
• Form/Format: PBS pH 7.2, 1% BSA, 0.05% NaN3
• Sequence Length: 373

• Applicable Applications for anti-CD14 antibody: Direct Immunofluorescence (IF), Indirect Immunofluorescence (IF), Flow Cytometry (FC/FACS)
• Application Notes: Direct Immunofluorescence (Staining Procedure); Fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparations.; ; Proposed staining procedure for whole blood in short:; - For each sample add 50 ul of EDTA anti-coagulated blood to a 3-5 ml tube; - Add 20 ul of the appropriate monoclonal antibody conjugate; - Incubate the tube for 15 minutes at 4 degree C or at room temperature in the dark; - Add 100 ul NM-LYSE to each tube and incubate for 10 minutes at room temperature - Add 3-4 ml of destilled water and vortex, incubate for 5-10; minutes at room temperature; - Centrifuge tube for 5 minutes at 300 g; - Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid; - Analyze immediately or store samples at 2-8 degree C in the dark and analyze within 24 hours; Proposed staining procedure for MNC in short:; - Carefully add 20 ul antibody conjugate and 50-100 ul MNC to the bottom of a tube; - Vortex at low speed for 1-2 seconds; - Incubate for 15-30 minutes at 2-8 degree C or at room temperature; - Centrifuge tubes for 5 minutes at 300 g; - Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge cells again for 5 minutes at 300 g; - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1 % formaldehyde and store them at 2-8 degree C in the dark. Analyze; fixed cells within 24 hours; ; Indirect Immunofluorescence (Staining Procedure); - Mix 20 ul purified antibody with 50 ul whole blood or MNC suspension; - Incubate for 15 minutes at 2-8 degree C; - Wash cells with phosphate buffered saline (PBS); - Add to cell pellet 20 ul of affinity purified, fluorochrome labeled F(ab')2 anti mouse Ig antibodies; - Incubate for 15 minutes at 2-8 degree C; - Wash cells with phosphate buffered saline (PBS) or proceed as described for direct staining
• CE Mark: CE
• Preparation and Storage: Monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody. For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8 degree C (DO NOT FREEZE!) and protected from prolonged exposure to light. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended.

Related Product Information for anti-CD14 antibody

CD14 is a GPI-anchored molecule expressed by virtually all human monocytes and macrophages and - to a lesser degree - granulocytes. CD14 together with Toll-like receptor 4 and MD-2 forms the LPS-receptor complex that recognizes and signals the presence of LPS. While CD14 has no signaling structure its main role seems to be the binding of LPS.

The MEM18 antibody permits the identification and enumeration of leukocytes using flow cytometry. MEM18 has been also used for functional studies since this antibody blocks the interaction of LPS with CD14 on monocytes. Results must be put within the context of other tests as well as the clinical history of the patient by a certified professional before final interpretation. Analysis performed with this antibody should be paralleled by positive and negative controls.

References

Beutler, B. (2002) Curr Top Microbiol Immunol 270, 109-20.
Goyert, S. M. (1989) In Leukocyte Typing IV (Oxford University Press, Oxford-New York-Tokyo) p789-793
Goyert, S. M., Ferrero, E., Rettig, W. J., Yenamandra, A. K., Obata, F. & Le Beau, M. M. (1988) Science 239, 497-500.
Goyert, S. M., Ferrero, E. M., Seremetis, S. V., Winchester, R. J., Silver, J. & Mattison, A. C. (1986) J Immunol 137, 3909-14.
Juan, T. S., Hailman, E., Kelley, M. J., Busse, L. A., Davy, E., Empig, C. J., Narhi, L. O., Wright S. D. & Lichenstein, H. S. (1995) J Biol Chem 270, 5219-24.
Knapp, W. (1989) In Leukocyte typing IV (Oxford University Press, Oxford-New York-Tokyo) p747-780
Means, T. K., Lien, E., Yoshimura, A., Wang, S., Golenbock, D. T. & Fenton, M. J. (1999) J Immunol 163, 6748-55.
Zilberman, M., Goyert, S. M. & Vogel, S. N. (2001) J Immunol 166, 574-81.
Tapping, R. I., Akashi, S., Miyake, K., Godowski, P. J. & Tobias, P. S. (2000) J Immunol 165, 5780-7.
Ugolini, V., Nunez, G., Smith, R. G., Stastny, P. & Capra, J. D. (1980) Proc Natl Acad Sci U S A 77, 6764-8.
Yoshimura, A., Lien, E., Ingalls, R. R., Tuomanen, E., Dziarski, R. & Golenbock, D. (1999) J Immunol 163, 1-5.

NCBI and Uniprot Product Information

• NCBI GI #: 2274810
• NCBI GeneID: 281048
• Molecular Weight: 39,667 Da
• NCBI Official Full Name: CD14
• NCBI Official Synonym Full Names: CD14 molecule<
• NCBI Official Symbol: CD14
• NCBI Protein Information: monocyte differentiation antigen CD14; CD14 antigen; myeloid cell-specific leucine-rich glycoprotein
• UniProt Protein Name: Monocyte differentiation antigen CD14
• Protein Family: Monocyte differentiation antigen
• UniProt Gene Name: CD14
• UniProt Entry Name: CD14_BOVIN

Uniprot Description

Coreceptor for bacterial lipopolysaccharide. In concert with LBP, binds to monomeric lipopolysaccharide and delivers it to the LY96/TLR4 complex, thereby mediating the innate immune response to bacterial lipopolysaccharide (LPS). Acts via MyD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Acts as a coreceptor 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. Binds electronegative LDL (LDL-) and mediates the cytokine release induced by LDL- ().