The antibody was purified by affinity chromatography, and conjugated with Alexa Fluor® 647 under optimal conditions. The solution is free of unconjugated Alexa Fluor® 647.
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Application:
FC
Recommended Usage:
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is 5 µl per million cells or 5 µl per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm. ** Alexa Fluor® is a registered trademark of Molecular Probes, Inc. Alexa Fluor® dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
Application Notes:
Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections, blocking of LPS stimulation4, and immunofluorescence microscopy5. The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for functional assays (Cat. No. 301810).
Application References:
1. McMichael, A., et al., 1987. Leucocyte Typing III. Oxford University Press. New York. 2. Knapp, W., et al., Eds. 1989. Leucocyte Typing IV. Oxford University Press. New York. 3. Schlossman, S., et al., Eds. 1995. Leucocyte Typing V. Oxford University Press. New York. 4. Power, C. P., et al., 2004. J. Immunol. 173:5229. 5. Williams, K. C., et al., 2001. J. Exp. Med. 193:905. 6. Iwamoto, S., et al., 2007. J. Immunol. 179:1449.
Human peripheral blood monocytes stained with M5E2 Alexa Fluor® 647
Description:
CD14 is a 53-55 kD glycosylphosphatidylinositol (GPI)-linked membrane glycoprotein also known as LPS receptor. CD14 is expressed at high levels on monocytes and macrophages, and at lower levels on granulocytes. Some dendritic cell populations such as interfollicular dendritic cells, reticular dendritic cells, and Langerhans cells have also been reported to express CD14. As a high-affinity receptor for LPS, CD14 is involved in the clearance of gram-negative pathogens and in the upregulation of adhesion molecules and cytokines expression in monocytes and neutrophils. The M5E2 antibody inhibits monocyte activation and cytokine production induced by LPS.
Other Names:
LPS receptor
Structure:
GPI-linked membrane glycoprotein, 53-55 kD
Distribution:
Monocytes, macrophages, granulocytes (low)
Function:
LPS receptor, clearance of Gram-negative pathogens
Ligand Receptor:
LPS
Antigen References:
1. Stocks, S., et al., 1990. Biochem. J. 268:275. 2. Wright, S., et al., 1990. Science 249:1434.
