Overview

  • Product name
    Anti-CD14 antibody [EPR3653]
    See all CD14 primary antibodies
  • Description
    Rabbit monoclonal [EPR3653] to CD14
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human CD14 aa 300-400. The exact sequence is proprietary.

  • Positive control
    • IHC-P: human colon, placenta and tonsil tissue; WB: Human tonsi, SW480 and PBMCl lysate.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133335 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
WB 1/1000 - 1/5000. Detects a band of approximately 53 kDa (predicted molecular weight: 40 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

For unpurified use at 1/500 - 1/1000.

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

  • Anti-CD14 antibody [EPR3653] (ab133335) at 1/1000 dilution (purified) + His-Tagged Human CD14 (aa20 to 345) recombinant protein at 0.015 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 40 kDa
    Observed band size: 43 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling CD14 with purified ab133335 at 1/2000 dilution (0.04 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
  • All lanes : Anti-CD14 antibody [EPR3653] (ab133335) at 1/1000 dilution (purified)

    Lane 1 : Human tonsil tissue lysate
    Lane 2 : HeLa (Human cervix adenocarcinoma) whole cell lysate
    Lane 3 : U-937 (Human histiocytic lymphoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
    Lanes 2-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 40 kDa
    Observed band size: 53 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking and diluting buffer: 5% NFDM/TBST

    The expression level in HeLa and U-937 are low (PMID: 9886426 and 15730927)

  • Immunohistochemical analysis of Formalin-fixed, paraffin- embedded human tonsil tissue labelling CD14 with ab133335 (unpurified) at 1/500 dilution. No blocking step performed. Anti-Rabbit HRP polymer was used as the secondary detection system. Heat-mediated antigen retrieval was performed using EDTA based pH 9.0 buffer.

  • Immunohistochemical analysis of Formalin-fixed, paraffin- embedded Human tonsil tissue labelling CD14 with ab133335 (unpurified) at 1/500 dilution.

  • Immunohistochemical analysis of Formalin-fixed, paraffin- embedded Human placenta tissue labelling CD14 with ab133335 (unpurified) at 1/500 dilution.

  • All lanes : Anti-CD14 antibody [EPR3653] (ab133335) at 1/1000 dilution (unpurified)

    Lane 1 : PBMC cell lysate
    Lane 2 : Human tonsil cell lysate
    Lane 3 : SW480 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 40 kDa
    Observed band size: 53 kDa why is the actual band size different from the predicted?

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Jamaly S  et al. Elevated plasma levels of P-selectin glycoprotein ligand-1-positive microvesicles in patients with unprovoked venous thromboembolism. J Thromb Haemost N/A:N/A (2018). Read more (PubMed: 29851269) »
  • Chakraborty B  et al. Deciphering genetic regulation of CD14 by SP1 through characterization of peripheral blood mononuclear transcriptome of P. faiciparum and P. vivax infected malaria patients. EBioMedicine 37:442-452 (2018). Read more (PubMed: 30337251) »
See all 7 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

Filter by Ratings

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer ph 8,5
Sample
Human Tissue sections (spleen)
Specification
spleen
Fixative
Paraformaldehyde

Mr. Rudolf Jung

Verified customer

Submitted Oct 22 2014

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