Overview

  • Product name

    Anti-CD14 antibody [EPR21847]
    See all CD14 primary antibodies
  • Description

    Rabbit monoclonal [EPR21847] to CD14
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, Flow Cyt, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse CD14 aa 1-350. The exact sequence is proprietary.
    Database link: P10810

  • Positive control

    • WB: J774A.1 and RAW264.7 whole cell lysate; mouse lymph node and placenta lysates. ICC/IF: J774A.1 and RAW 264.7 cells. Flow cyt: RAW 264.7 cells. IP: RAW 264.7 whole cell lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab221678 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP 1/30.
Flow Cyt 1/500.
ICC/IF 1/1000.
WB 1/1000. Detects a band of approximately 50-55 kDa (predicted molecular weight: 39 kDa).

Target

  • Function

    Cooperates with MD-2 and TLR4 to mediate 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. Up-regulates cell surface molecules, including adhesion molecules.
  • Tissue specificity

    Expressed strongly on the surface of monocytes and weakly on the surface of granulocytes; also expressed by most tissue macrophages.
  • Sequence similarities

    Contains 11 LRR (leucine-rich) repeats.
  • Post-translational
    modifications

    N- and O- glycosylated. O-glycosylated with a core 1 or possibly core 8 glycan.
  • Cellular localization

    Cell membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • CD 14 antibody
    • CD_antigen=CD14 antibody
    • CD14 antibody
    • CD14 antigen antibody
    • CD14 molecule antibody
    • CD14_HUMAN antibody
    • LPS-R antibody
    • Mo2 antibody
    • Monocyte differentiation antigen CD14 antibody
    • Monocyte differentiation antigen CD14 urinary form antibody
    • Monocyte differentiation antigen CD14, membrane-bound form antibody
    • Myeloid cell specific leucine rich glycoprotein antibody
    • Myeloid cell-specific leucine-rich glycoprotein antibody
    see all

Images

  • Flow cytometric analysis of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell line labeling CD14 with ab221678 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)(ab150077) at 1/2000 dilution was used as the secondary antibody.

    Gated on viable cells.

  • CD14 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab221678 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221678 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

    Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg (Input).
    Lane 2: ab221678 IP in RAW 264.7 whole cell lysate(+).
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab221678 in RAW 264.7 whole cell lysate (-).

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 8 seconds

    The molecular mass observed is consistent with the literature (PMID: 9502426; PMID:7513013)

  • All lanes : Anti-CD14 antibody [EPR21847] (ab221678) at 1/1000 dilution

    Lane 1 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate at 10 µg
    Lane 2 : Mouse lymph node lysate at 20 µg
    Lane 3 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg
    Lane 4 : Mouse placenta lysate at 10 µg

    Secondary
    Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Developed using the ECL technique.

    Predicted band size: 39 kDa
    Observed band size: 50-55 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: Lane 1: 6 seconds; Lane 2: 3 minutes; Lane 3: 10 seconds; Lane 4: 81 seconds.

    The molecular mass observed is consistent with the literature (PMID: 9502426; PMID: 7513013).

  • Immunofluorescent analysis of 100% methanol-fixed RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling CD14 with ab221678 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)(ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on RAW 264.7 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (re

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)(ab150077) secondary at 1/1000 dilution.

     

  • Immunofluorescent analysis of 100% methanol-fixed J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) cells labeling CD14 with ab221678 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on J774A.1 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.

     

     

References

ab221678 has not yet been referenced specifically in any publications.

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