Recombinant
RabMAb

Recombinant Anti-CD14 antibody [EPR3653] (ab133335)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3653] to CD14
  • Suitable for: WB, IHC-P
  • Reacts with: Human

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
    Unsuitable for: ICC/IF

  • 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 tonsil tissue lysate, SW480 and PBMCl lysates.

  • 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.

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

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.
  • Application notes
    Is unsuitable for ICC/IF.

    • 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

      • Western blot - Anti-CD14 antibody [EPR3653] (ab133335)
        Western blot - Anti-CD14 antibody [EPR3653] (ab133335)
        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) - Anti-CD14 antibody [EPR3653] (ab133335)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)

        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 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.

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

        Lane 1 : Human tonsil lysates prepared in RIPA lysis method
        Lane 2 : Human tonsil lysates prepared in 1%SDS Hot lysis method

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

        Predicted band size: 40 kDa


        Exposure time: 45 seconds


        Blocking/Diluting buffer and concentration: 5% NFDM/TBST

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

        Lane 1 : Human tonsil tissue lysate prepared in 1% SDS Hot lysis method
        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)

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)

        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.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)

        Immunohistochemical analysis of Formalin-fixed, paraffin- embedded Human tonsil tissue labelling CD14 with ab133335 (unpurified) at 1/500 dilution. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)

        Immunohistochemical analysis of Formalin-fixed, paraffin- embedded Human placenta tissue labelling CD14 with ab133335 (unpurified) at 1/500 dilution. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

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

        Lane 1 : PBMC cell lysate prepared in 1% SDS Hot lysis method
        Lane 2 : Human tonsil tissue lysate prepared in 1%SDS Hot lysis method
        Lane 3 : SW480 cell lysate prepared in 1%SDS Hot lysis method

        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?

      • Other - Anti-CD14 antibody [EPR3653] (ab133335)
        Other - Anti-CD14 antibody [EPR3653] (ab133335)
        Equilibrium disassociation constant (KD)
        Learn more about KD

        Click here to learn more about KD

      References

      Publishing research using ab133335? Please let us know so that we can cite the reference in this datasheet.

      ab133335 has been referenced in 10 publications.

      • Feng Y  et al. LncRNA NR-104098 Inhibits AML Proliferation and Induces Differentiation Through Repressing EZH2 Transcription by Interacting With E2F1. Front Cell Dev Biol 8:142 (2020). PubMed: 32296698
      • Wheeler MA  et al. Environmental Control of Astrocyte Pathogenic Activities in CNS Inflammation. Cell 176:581-596.e18 (2019). PubMed: 30661753
      • 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). 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). PubMed: 30337251
      • Gao N  et al. ß2-Microglobulin participates in development of lung emphysema by inducing lung epithelial cell senescence. Am J Physiol Lung Cell Mol Physiol 312:L669-L677 (2017). PubMed: 28213472
      • Chen J  et al. The pathological structure of the perivascular niche in different microvascular patterns of glioblastoma. PLoS One 12:e0182183 (2017). PubMed: 28771552
      • Perelygina L  et al. Rubella persistence in epidermal keratinocytes and granuloma M2 macrophages in patients with primary immunodeficiencies. J Allergy Clin Immunol 138:1436-1439.e11 (2016). IF ; Human . PubMed: 27613149
      • Dutertre CA  et al. Deciphering the stromal and hematopoietic cell network of the adventitia from non-aneurysmal and aneurysmal human aorta. PLoS One 9:e89983 (2014). IF, IHC ; Human . PubMed: 24587165
      • Hsu RY  et al. LPS-induced TLR4 signaling in human colorectal cancer cells increases beta1 integrin-mediated cell adhesion and liver metastasis. Cancer Res 71:1989-98 (2011). PubMed: 21363926
      • Kayamori K  et al. Roles of interleukin-6 and parathyroid hormone-related peptide in osteoclast formation associated with oral cancers: significance of interleukin-6 synthesized by stromal cells in response to cancer cells. Am J Pathol 176:968-80 (2010). PubMed: 20035059

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      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD14 antibody [EPR3653]

       Good
      Abreviews
      abreview image
      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
      Read More

      Mr. Rudolf Jung

      Verified customer

      Submitted Oct 22 2014

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