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RecombinantRabMAb

Recombinant Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

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Flow Cytometry - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
  • Immunohistochemistry (Frozen sections) - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
  • Flow Cytometry - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
  • Immunoprecipitation - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
  • Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

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 - BSA and Azide free
  • Suitable for: IHC-Fr, WB, ICC/IF, IP, Flow Cyt
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 Unconjugated

Overview

  • Product name

    Anti-CD14 antibody [EPR21847] - BSA and Azide free
    See all CD14 primary antibodies

  • Description

    Rabbit monoclonal [EPR21847] to CD14 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-Fr, WB, ICC/IF, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse

  • Immunogen

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

  • Positive control

    • ICC/IF: J774A.1 cells; IHC-Fr: mouse spleen tissue; Flow Cyt: RAW 264.7 C57 BL/6 mouse bone marrow cells; IP: RAW 264.7.

  • General notes

    ab231852 is the carrier-free version of ab221678.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab231852 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.
WB
Use at an assay dependent concentration. Detects a band of approximately 50-55 kDa (predicted molecular weight: 39 kDa).
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.
Notes
IHC-Fr
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 50-55 kDa (predicted molecular weight: 39 kDa).
ICC/IF
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

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] - BSA and Azide free (ab231852)
      Flow Cytometry - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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] - BSA and Azide free (ab231852)
      Immunohistochemistry (Frozen sections) - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

      This data was developed using ab221678 ,the same antibody clone in a different buffer formulation.

      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] - BSA and Azide free (ab231852)
      Flow Cytometry - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

    • Immunoprecipitation - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
      Immunoprecipitation - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

      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)

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

    • Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
      Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

      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.

       

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

    • Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
      Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

    • Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)
      Anti-CD14 antibody [EPR21847] - BSA and Azide free (ab231852)

    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 (0)

    Publishing research using ab231852? Please let us know so that we can cite the reference in this datasheet.

    ab231852 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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