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RecombinantRabMAb

Recombinant Anti-CD14 antibody [EPR3653] (ab133335)

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

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

Conjugates logo Related conjugates and formulations

Carrier Free HRP

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Primary
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Anti-SSEA1 antibody [MC-480] (ab16285)
Secondary
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Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Protein
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Recombinant Mouse CD14 protein (ab207103)

View more associated products

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

  • Positive control

    • IHC-P: human colon, placenta and tonsil tissue; WB: Human tonsil tissue lysate, SW480 and PBMCl lysates.
  • 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.

    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.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Dissociation constant (KD)

    KD = 4.70 x 10 -12 M
    Learn more about KD
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3653
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Cell Type Markers
    • CD
    • Myeloid Cells
    • Stem Cells
    • Hematopoietic Progenitors
    • Hematopoietic Stem Cells
    • Human Lineage Negative
    • Stem Cells
    • Mesenchymal Stem Cells
    • Negative Markers
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Monocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Neutrophil Lineage
    • Immunology
    • Innate Immunity
    • TLR Signaling
    • Stem Cells
    • Endothelial Progenitors
    • Endothelial Markers
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • CD markers ELISA kits

Associated products

  • Alternative Versions

    • HRP Anti-CD14 antibody [EPR3653] (ab195525)
    • Anti-CD14 antibody [EPR3653] - Low endotoxin, Azide free (ab214438)
    • Anti-CD14 antibody [EPR3653] - BSA and Azide free (ab226121)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • SW480 whole cell lysate (ab3957)
  • Recombinant Protein

    • Recombinant Mouse CD14 protein (ab207103)

Applications

The Abpromise guarantee

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)
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.

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.
  • Target information above from: UniProt accession P08571 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 929 Human
    • Omim: 158120 Human
    • SwissProt: P08571 Human
    • Unigene: 163867 Human
    • 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

    • 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)
      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

    • 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?

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

      Click here to learn more about KD
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (ab133335)

      Tissue Microarrays stained for "Anti-CD14 antibody [EPR3653]” using "ab133335" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab133335 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Anti-CD14 antibody [EPR3653] (ab133335)
      Anti-CD14 antibody [EPR3653] (ab133335)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (14)

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

    ab133335 has been referenced in 14 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
    • Chang HF  et al. Investigation of Cytotoxic T Lymphocyte Function during Allorejection in the Anterior Chamber of the Eye. Int J Mol Sci 21:N/A (2020). PubMed: 32629968
    • Hasegawa T  et al. Reduction in Human Epidermal Langerhans Cells with Age Is Associated with Decline in CXCL14-Mediated Recruitment of CD14+ Monocytes. J Invest Dermatol 140:1327-1334 (2020). PubMed: 31881212
    • Aguirre LA  et al. Tumor stem cells fuse with monocytes to form highly invasive tumor-hybrid cells. Oncoimmunology 9:1773204 (2020). PubMed: 32923132
    • Feng Y  et al. Long noncoding RNA HOXA-AS2 functions as an oncogene by binding to EZH2 and suppressing LATS2 in acute myeloid leukemia (AML). Cell Death Dis 11:1025 (2020). PubMed: 33268767
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

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

    Good
    Abreviews
    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
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Rudolf Jung

    Verified customer

    Submitted Oct 22 2014

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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