Skip to main content Skip to navigation
Hamburger Menu
Search icon
Country Selector Country Selector
China (Chinese)
Profile Icon Profile Icon
登录/注册 UserName
产品

解决方案

发现&学习

资源&工具

Support

Search icon Search icon
Close Menu
      Alert Icon
      PerCP-Cy™5.5 Rat Anti-Mouse CD14
      PerCP-Cy™5.5 Rat Anti-Mouse CD14
      Flow cytometric analysis of CD14 on J774A.1 cells.  J774A.1 cells (Mouse monocyte/macrophage cells; ATCC TIB-67) were stained either with a PerCP-Cy™5.5 Rat IgG1, κ isotype control (shaded) or with the PerCP-Cy™5.5 Rat Anti-Mouse CD14 antibody (unshaded).  Histograms were derived from gated events based on light scattering characteristics for J774A.1 cells.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.
      PerCP-Cy™5.5 Rat Anti-Mouse CD14
      Flow cytometric analysis of CD14 on J774A.1 cells.  J774A.1 cells (Mouse monocyte/macrophage cells; ATCC TIB-67) were stained either with a PerCP-Cy™5.5 Rat IgG1, κ isotype control (shaded) or with the PerCP-Cy™5.5 Rat Anti-Mouse CD14 antibody (unshaded).  Histograms were derived from gated events based on light scattering characteristics for J774A.1 cells.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.
      商品详情
      Down Arrow Up Arrow


      BD Pharmingen™
      Mouse (QC Testing)
      Rat LOU, also known as Louvain, LOU/C, LOU/M IgG1, κ
      Recombinant Mouse CD14
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      12475
      AB_1727428
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      准备和存储

      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 with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
      显示较多信息 blue down arrow 显示较少信息 blue up arrow

      推荐的实验流程

      Flow Cytometry:  Investigators should note that Mouse BD Fc Block™ purified rat anti-CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142) and antibodies of the rat IgG2b isotype may potentially interfere with the reactivity of the PerCP-Cy™5.5 Rat Anti-Mouse CD14 antibody (clone rmC5-3) in a concentration-dependent manner. For alternative methods for inhibition of non-specific reactivity, investigators may find the use of purified mouse IgG at a 10-100-fold excess to be more appropriate.

      显示较多信息 blue down arrow 显示较少信息 blue up arrow

      商品通知

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
      4. This PerCP-conjugated product is sold under license to the following patent: US Patent No. 4,876,190.
      5. Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
      6. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
      7. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
      8. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
      9. 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.
      10. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      显示较多信息 blue down arrow 显示较少信息 blue up arrow
      560638 Rev. 1
      抗体详情
      Down Arrow Up Arrow
      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.

      560638 Rev. 1
      格式详情
      Down Arrow Up Arrow
      PerCP-Cy5.5
      PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PerCP-Cy5.5
      Blue 488 nm
      482 nm
      676 nm
      560638 Rev.1
      报价单和参考
      Down Arrow Up Arrow

      研发参考 (20)

      1. Adachi Y, Satokawa C, Saeki M, et al. Inhibition by a CD14 monoclonal antibody of lipopolysaccharide binding to murine macrophages. J Endotoxin Res. 1999; 5(3):139-146. (Biology).
      2. Akashi S, Saitoh S, Wakabayashi Y, et al. Lipopolysaccharide interaction with cell surface Toll-like receptor 4-MD-2: higher affinity than that with MD-2 or CD14. J Exp Med. 2003; 198(7):1035-1042. (Biology). 查看参考
      3. 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). 查看参考
      4. 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). 查看参考
      5. 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). 查看参考
      6. 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. (Biology). 查看参考
      7. Heumann D, Adachi Y, Le Roy D, et al. Role of plasma, lipopolysaccharide-binding protein, and CD14 in response of mouse peritoneal exudate macrophages to endotoxin. Infect Immun. 2001; 69(1):378-385. (Biology). 查看参考
      8. Le Roy D, Di Padova F, Adachi Y, Glauser MP, Calandra T, Heumann D. Critical role of lipopolysaccharide-binding protein and CD14 in immune responses against gram-negative bacteria. J Immunol. 2001; 167(5):2759-2765. (Biology). 查看参考
      9. 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.
      10. 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: Western blot). 查看参考
      11. 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. (Biology). 查看参考
      12. Nagaoka I, Hirota S, Niyonsaba F, et al. Cathelicidin family of antibacterial peptides CAP18 and CAP11 inhibit the expression of TNF-alpha by blocking the binding of LPS to CD14(+) cells. J Immunol. 2001; 167(6):3329-3338. (Biology). 查看参考
      13. 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). 查看参考
      14. 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. (Biology). 查看参考
      15. 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. (Biology). 查看参考
      16. 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. (Biology). 查看参考
      17. Tasaka S, Ishizaka A, Yamada W, et al. Effect of CD14 blockade on endotoxin-induced acute lung injury in mice. Am J Respir Cell Mol Biol. 2003; 29(2):252-258. (Biology). 查看参考
      18. Triantafilou M, Triantafilou K. Lipopolysaccharide recognition: CD14, TLRs and the LPS-activation cluster. Trends Immunol. 2002; 23(6):301-304. (Biology). 查看参考
      19. Yang S, Sugawara S, Monodane T, et al. Micrococcus luteus teichuronic acids activate human and murine monocytic cells in a CD14- and toll-like receptor 4-dependent manner. Infect Immun. 2001; 69(4):2025-2030. (Biology). 查看参考
      20. Ziegler-Heitbrock HW. Heterogeneity of human blood monocytes: the CD14+ CD16+ subpopulation. Immunol Today. 1996; 17(9):424-428. (Biology). 查看参考
      查看所有文件 (20) 查看更少内容
      560638 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.

      Website Feedback