Mouse anti CD14, conjugated to FITC

Mouse anti CD14, conjugated to FITC

• Background: 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 interpreted by a certified professional before final interpretation. Analyses performed with this antibody should be paralleled by positive and negative controls. If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us
• CAS Number: 9007-83-4
• UniProt: P08572
• Host: Mouse
• Species Reactivity: Human
• Isotype: IgG1
• Clone: MEM18
• Conjugation: FITC
• Type: Primary Antibodies
• Applications: Direct IF, IF, Flow Cytometry
• Assay Principle: Direct Immunofluorescence (Staining Procedure) Nordic-MUbio 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 µL of EDTA anti-coagulated blood to a 3-5 mL tube - Add 20 µL of the appropriate Nordic-MUbio monoclonal antibody conjµgate - Incubate the tube for 15 minutes at 4°C or at room temperature in the dark - Add 100 µL NM-LYSE (Cat.No. GAS-003) 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 - Centrifµge 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° C in the dark and analyze within 24 hours For “No-Wash” protocol please refer to www.nordicmubio.com Proposed staining procedure for MNC in short: - Carefully add 20 µL antibody conjµgate and 50-100 µL MNC to the bottom of a tube - Vortex at low speed for 1-2 seconds - Incubate for 15-30 minutes at 2-8°C or at room temperature - Centrifµge tubes for 5 minutes at 300 g - Remove supernatant, resuspend cells in 2-5 mL of phosphate buffered saline (PBS) and centrifµge 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°C in the dark. Analyze fixed cells within 24 hours Indirect Immunofluorescence (Staining Procedure) - Mix 20 µL Nordic-MUbio purified antibody with 50 µL whole blood or MNC suspension - Incubate for 15 minutes at 2-8°C - Wash cells with phosphate buffered saline (PBS) - Add to cell pellet 20 µL of affinity purified, fluorochrome labeled F(ab’)2 anti mouse Ig antibodies - Incubate for 15 minutes at 2-8°C - Wash cells with phosphate buffered saline (PBS) or proceed as described for direct staining
• Form: 2 mL of FITC-conjµgated anti CD14 (clone MEM18) in PBS pH 7,2, 1% BSA, and 0,05% NaN3, approximately 100 tests
• Precautions: For professional users only. This reagent contains sodium azide. To avoid the development of hazardous conditions, reagents containing azide should be diluted in running water prior to be discarded. Similar to the work with other biological products, proper handling procedures are recommended.
• References & Citations: 1. Beutler, B. (2002) Curr Top Microbiol Immunol 270, 109-20. _x000D_ 2. Goyert, S. M. (1989) In Leukocyte Typing IV (Oxford University Press, Oxford-New York-Tokyo) p789-793_x000D_ 3. Goyert, S. M., Ferrero, E., Rettig, W. J., Yenamandra, A. K., Obata, F. , Le Beau, M. M. (1988) Science 239, 497-500. _x000D_ 4. Goyert, S. M., Ferrero, E. M., Seremetis, S. V., Winchester, R. J., Silver, J. , Mattison, A. C. (1986) J Immunol 137, 3909-14. _x000D_ 5. 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. _x000D_ 6. Knapp, W. (1989) In Leukocyte typing IV (Oxford University Press, Oxford-New York-Tokyo) p747-780 _x000D_ 7. Means, T. K., Lien, E., Yoshimura, A., Wang, S., Golenbock, D. T. , Fenton, M. J. (1999) J Immunol 163, 6748-55. _x000D_ 8. Zilberman, M., Goyert, S. M. , Vogel, S. N. (2001) J Immunol 166, 574-81. _x000D_ 9. Tapping, R. I., Akashi, S., Miyake, K., Godowski, P. J. , Tobias, P. S. (2000) J Immunol 165, 5780-7. _x000D_ 10. Ugolini, V., Nunez, G., Smith, R. G., Stastny, P. , Capra, J. D. (1980) Proc Natl Acad Sci U S A 77, 6764-8. _x000D_ 11. Yoshimura, A., Lien, E., Ingalls, R. R., Tuomanen, E., Dziarski, R. , Golenbock, D. (1999) J Immunol 163, 1-5.
• Storage Conditions: Nordic-MUbio 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°C (DO NOT FREEZE!) and protected from prolonged exposure to light. If a slight precipitation occurs upon storage, this should be removed by centrifugation; It will not affect the performance or the concentration of the product 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.