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RecombinantRabMAb

Recombinant Anti-CD14 antibody [EPR21847] (ab221678)

Submit a review Submit a question References (8)
Flow Cytometry - Anti-CD14 antibody [EPR21847] (ab221678)
  • Immunohistochemistry (Frozen sections) - Anti-CD14 antibody [EPR21847] (ab221678)
  • Flow Cytometry - Anti-CD14 antibody [EPR21847] (ab221678)
  • Immunoprecipitation - Anti-CD14 antibody [EPR21847] (ab221678)
  • Western blot - Anti-CD14 antibody [EPR21847] (ab221678)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] (ab221678)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] (ab221678)
  • Anti-CD14 antibody [EPR21847] (ab221678)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21847] to CD14
  • Suitable for: IHC-Fr, IP, Flow Cyt, ICC/IF, WB
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

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: IHC-Fr, IP, Flow Cyt, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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, C57 BL/6 mouse bone marrow cells. IP: RAW 264.7 whole cell lysate; IHC-Fr: Mouse spleen tissue.

  • General notes

    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.

Properties

Applications

The Abpromise guarantee

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
IHC-Fr
Use at an assay dependent concentration.
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).
Notes
IHC-Fr
Use at an assay dependent concentration.
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 Cytometry - Anti-CD14 antibody [EPR21847] (ab221678)
      Flow Cytometry - Anti-CD14 antibody [EPR21847] (ab221678)

      Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab221678 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab221678 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 µl at 10.0 μg/ml (1/215)) for 30min on ice. The cells were simultaneously stained with Ly6G.

      The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

      Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

    • Immunohistochemistry (Frozen sections) - Anti-CD14 antibody [EPR21847] (ab221678)
      Immunohistochemistry (Frozen sections) - Anti-CD14 antibody [EPR21847] (ab221678)

      Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen tissue labeling CD14 with ab221678 at 1/50 (10.62 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) at 1/1000 dilution (Green). Positive staining on mouse spleen. is observed. The nuclear counterstain was DAPI (Blue).

      Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor ®; 488) at 1000 dilution.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

    • Flow Cytometry - Anti-CD14 antibody [EPR21847] (ab221678)
      Flow Cytometry - Anti-CD14 antibody [EPR21847] (ab221678)

      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.

    • Immunoprecipitation - Anti-CD14 antibody [EPR21847] (ab221678)
      Immunoprecipitation - Anti-CD14 antibody [EPR21847] (ab221678)

      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)

    • Western blot - Anti-CD14 antibody [EPR21847] (ab221678)
      Western blot - Anti-CD14 antibody [EPR21847] (ab221678)
      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).

    • Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] (ab221678)
      Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] (ab221678)

      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.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] (ab221678)
      Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] (ab221678)

      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.

       

       

    • Anti-CD14 antibody [EPR21847] (ab221678)
      Anti-CD14 antibody [EPR21847] (ab221678)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (8)

    Publishing research using ab221678? Please let us know so that we can cite the reference in this datasheet.

    ab221678 has been referenced in 8 publications.

    • Zhao L  et al. Dendritic cell-mediated chronic low-grade inflammation is regulated by the RAGE-TLR4-PKCβ1 signaling pathway in diabetic atherosclerosis. Mol Med 28:4 (2022). PubMed: 35062863
    • Pei S  et al. Lysophosphatidic Acid Receptor 3 Suppress Neutrophil Extracellular Traps Production and Thrombosis During Sepsis. Front Immunol 13:844781 (2022). PubMed: 35464399
    • Lin C  et al. PTEN-induced kinase 1 enhances the reparative effects of bone marrow mesenchymal stromal cells on mice with renal ischaemia/reperfusion-induced acute kidney injury. Hum Cell 35:1650-1670 (2022). PubMed: 35962179
    • Yang R  et al. Telocytes-derived extracellular vesicles alleviate aortic valve calcification by carrying miR-30b. ESC Heart Fail 8:3935-3946 (2021). PubMed: 34165260
    • Li Y  et al. Polarization of rheumatoid macrophages is regulated by the CDKN2B-AS1/ MIR497/TXNIP axis. Immunol Lett 239:23-31 (2021). PubMed: 34418490
    • Woo J  et al. High-throughput and high-efficiency sample preparation for single-cell proteomics using a nested nanowell chip. Nat Commun 12:6246 (2021). PubMed: 34716329
    • Rao L  et al. Hybrid cellular membrane nanovesicles amplify macrophage immune responses against cancer recurrence and metastasis. Nat Commun 11:4909 (2020). PubMed: 32999291
    • Sun Q  et al. Bactericidal/Permeability-Increasing Protein Improves Cognitive Impairment in Diabetic Mice via Blockade of the LPS-LBP-TLR4 Signaling Pathway. Front Physiol 11:718 (2020). PubMed: 33643054

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