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      Alexa Fluor® 647 Rat Anti-Mouse CD14
      Alexa Fluor® 647 Rat Anti-Mouse CD14
      Flow cytometric analysis of CD14 expression on mouse J774A.1 cells. Cells from the J774A.1 (Mouse macrophage, ATCC TIB-67) cell line were stained with either Alexa Fluor® 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; dashed line histogram) or Alexa Fluor® 647 Rat Anti-Mouse CD14 antibody (Cat. No. 565743; solid line histogram). The fluorescence histogram showing CD14 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable J774A cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      Alexa Fluor® 647 Rat Anti-Mouse CD14

      Immunohistofluorescent analysis of CD14 expression by cells within C57BL/6 mouse spleen. A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with Alexa Fluor® 647 Rat Anti-Mouse CD14 antibody (Cat. No. 565743, pseudocolored green), BD Horizon™ BV421 Rat Anti-Mouse CD3 Molecular Complex antibody (Cat. No. 564008, pseudocolored red), BD Horizon™ BV480 Rat Anti-Mouse CD31 antibody (Cat. No. 565629, pseudocolored cyan), and Alexa Fluor® 488 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557669, pseudocolored blue). Images were acquired and merged using an epifluorescence microscopic system. Original magnification, 20x.

      Alexa Fluor® 647 Rat Anti-Mouse CD14 Alexa Fluor® 647 Rat Anti-Mouse CD14
      Flow cytometric analysis of CD14 expression on mouse J774A.1 cells. Cells from the J774A.1 (Mouse macrophage, ATCC TIB-67) cell line were stained with either Alexa Fluor® 647 Rat IgG1, κ Isotype Control (Cat. No. 557731; dashed line histogram) or Alexa Fluor® 647 Rat Anti-Mouse CD14 antibody (Cat. No. 565743; solid line histogram). The fluorescence histogram showing CD14 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable J774A cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

      Immunohistofluorescent analysis of CD14 expression by cells within C57BL/6 mouse spleen. A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with Alexa Fluor® 647 Rat Anti-Mouse CD14 antibody (Cat. No. 565743, pseudocolored green), BD Horizon™ BV421 Rat Anti-Mouse CD3 Molecular Complex antibody (Cat. No. 564008, pseudocolored red), BD Horizon™ BV480 Rat Anti-Mouse CD31 antibody (Cat. No. 565629, pseudocolored cyan), and Alexa Fluor® 488 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557669, pseudocolored blue). Images were acquired and merged using an epifluorescence microscopic system. Original magnification, 20x.

      Product Details
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      BD Pharmingen™
      Cd14; CD14 antigen; Myeloid cell-specific leucine-rich glycoprotein
      Mouse (QC Testing)
      Rat LOU, also known as Louvain, LOU/C, LOU/M IgG1, κ
      Recombinant Mouse CD14
      Flow cytometry (Routinely Tested), Immunofluorescence, Immunohistochemistry (Tested During Development)
      0.2 mg/ml
      12475
      AB_2739340
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
      4. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      6. 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.
      7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      8. An isotype control should be used at the same concentration as the antibody of interest.
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      565743 Rev. 1
      Antibody Details
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      rmC5-3

      The rmC5-3 monoclonal antibody specifically binds to residues 308-322 of the hydrophilic region of mouse CD14. CD14 is a 53-55 kDa glycophosphatidylinositol (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). CD14 serves as a receptor for LPS that can play a role in the cellular production of proinflammatory cytokines such as IL-1 and TNF. CD14 can be 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. 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.

      565743 Rev. 1
      Format Details
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      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      565743 Rev.1
      Citations & References
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      Development References (13)

      1. Fearns C, Kravchenko VV, Ulevitch RJ, Loskutoff DJ. Murine CD14 gene expression in vivo: extramyeloid synthesis and regulation by lipopolysaccharide. J Exp Med. 1995; 181(3):857-866. (Biology). View Reference
      2. Fearns C, Loskutoff DJ. Role of tumor necrosis factor alpha in induction of murine CD14 gene expression by lipopolysaccharide. Infect Immun. 1997; 65(11):4822-4831. (Biology). View Reference
      3. Ferrero E, Hsieh CL, Francke U, Goyert SM. CD14 is a member of the family of leucine-rich proteins and is encoded by a gene syntenic with multiple receptor genes. J Immunol. 1990; 145(1):331-336. (Biology). View Reference
      4. Haziot A, Ferrero E, Kontgen F, et al. Resistance to endotoxin shock and reduced dissemination of gram-negative bacteria in CD14-deficient mice. Immunity. 1996; 4(4):407-414. (Clone-specific: Flow cytometry). View Reference
      5. Mahnke K, Becher P, Ricciardi-Castagnoli P, Luger TA, Schawrz T Grabbe S. CD14 is expressed by subsets of murine dendritic cells and upregulated by lipopolysaccharide. In: Ricciardi-Castagnoli P, ed. Dendritic Cells in Fundamental and Clinical Immunology. New York: Plenum Press; 1997:145-159.
      6. Matsuura K, Ishida T, Setoguchi M, Higuchi Y, Akizuki S, Yamamoto S. Upregulation of mouse CD14 expression in Kupffer cells by lipopolysaccharide. J Exp Med. 1994; 179(5):1671-1676. (Immunogen: ELISA, Flow cytometry, Immunohistochemistry, Western blot). View Reference
      7. Miyata Y, Takeda H, Kitano S, Hanazawa S. Porphyromonas gingivalis lipopolysaccharide-stimulated bone resorption via CD14 is inhibited by broad-spectrum antibiotics. Infect Immun. 1997; 65(9):3513-3519. (Clone-specific: Immunohistochemistry). View Reference
      8. Nasu N, Yoshida S, Akizuki S, Higuchi Y, Setoguchi M, Yamamoto S. Molecular and physiological properties of murine CD14. Int Immunol. 1991; 3(2):205-213. (Biology). View Reference
      9. Pulendran B, Lingappa J, Kennedy MK, et al. Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice. J Immunol. 1997; 159(5):2222-2231. (Clone-specific: Flow cytometry). View Reference
      10. Stewart CC. Methods for studying the ontogeny of monnuclear phagocytes. In: Weir DM, Herzenberg LA, Blackwell C, ed. Weir's Handbook of Experiemental Immunology. Blackwell Science Publications; 1986:44.1-44.17.
      11. Takakuwa T, Knopf HP, Sing A, Carsetti R, Galanos C, Freudenberg MA. Induction of CD14 expression in Lpsn, Lpsd and tumor necrosis factor receptor-deficient mice. Eur J Immunol. 1996; 26(11):2686-2692. (Clone-specific: Flow cytometry). View Reference
      12. Takamatsu S, Nakashima I, Nakano K. Modulation of endotoxin-induced histamine synthesis by cytokines in mouse bone marrow-derived macrophages. J Immunol. 1996; 156(2):778-785. (Clone-specific: Flow cytometry). View Reference
      13. Takezawa R, Watanabe Y, Akaike T. Direct evidence of macrophage differentiation from bone marrow cells in the liver: a possible origin of Kupffer cells. J Biochem (Tokyo). 1995; 118(6):1175-1183. (Biology). View Reference
      View All (13) View Less
      565743 Rev. 1

      Please refer to Support Documents for Quality Certificates


      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


      Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

      For Research Use Only. Not for use in diagnostic or therapeutic procedures.

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