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Rabbit Monoclonal CD14 antibody. Suitable for IHC-P, WB and reacts with Human samples. Cited in 27 publications.


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (AB133335), expandable thumbnail
  • Western blot - Anti-CD14 antibody [EPR3653] (AB133335), expandable thumbnail
  • Western blot - Anti-CD14 antibody [EPR3653] (AB133335), expandable thumbnail
  • Western blot - Anti-CD14 antibody [EPR3653] (AB133335), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (AB133335), expandable thumbnail

Publications

  • Cancer immunology, immunotherapy : CII 72:4385-43972023
    Spatial immune composition of tumor microenvironment in patients with pancreatic cancer.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Eline S Zwart,Thomas van Ee,Alsya J Affandi,Lenka N C Boyd,Ernesto Rodriguez,Joke M M den Haan,Arantza Farina,Nicole C T van Grieken,Laura L Meijer,Yvette van Kooyk,Reina E Mebius,Geert Kazemier
    PubMed 37938368
  • Journal of cancer research and clinical oncology 149:14731-147432023
    The immune microenvironment after neoadjuvant therapy compared to upfront surgery in patients with pancreatic cancer.
    Applications:
    Unspecified application
    Reactive species:
    Unspecified reactive species
    Eline S Zwart,Thomas van Ee,Deesje Doppenberg,Arantza Farina,Johanna W Wilmink,Eva Versteijne,Olivier R Busch,Marc G Besselink,Laura L Meijer,Yvette van Kooyk,Reina E Mebius,Geert Kazemier
    PubMed 37587309

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IF
Human
Tested
Tested
Not recommended

Tested
Tested

Species

Human

Dilution info

1/2000

Notes

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

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

1/1000 - 1/5000

Notes

-

Not recommended
Not recommended

Species

Human

Dilution info

-

Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

3 products for Alternative Version

Target data

Function

Coreceptor for bacterial lipopolysaccharide (PubMed:1698311, PubMed:23264655). In concert with LBP, binds to monomeric lipopolysaccharide and delivers it to the LY96/TLR4 complex, thereby mediating the innate immune response to bacterial lipopolysaccharide (LPS) (PubMed:20133493, PubMed:23264655, PubMed:22265692). Acts via MyD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response (PubMed:8612135). Acts as a coreceptor for TLR2:TLR6 heterodimer in response to diacylated lipopeptides and for TLR2:TLR1 heterodimer in response to triacylated lipopeptides, these clusters trigger signaling from the cell surface and subsequently are targeted to the Golgi in a lipid-raft dependent pathway (PubMed:16880211). Binds electronegative LDL (LDL(-)) and mediates the cytokine release induced by LDL(-) (PubMed:23880187).

Alternative names

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Rabbit Monoclonal CD14 antibody. Suitable for IHC-P, WB and reacts with Human samples. Cited in 27 publications.

Alternative names

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR3653

Purification technique

Affinity purification Protein A

Dissociation constant

4.7 x 10-12 M

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

Activity summary

CD14 also known as cluster of differentiation 14 is a glycoprotein with a molecular mass of approximately 55 kDa. This protein is expressed mainly on the surface of macrophages monocytes and to a lesser extent on neutrophils. CD14 exists in two forms: membrane-bound (mCD14) and soluble (sCD14). It functions as a co-receptor along with TLR4 (Toll-like receptor 4) and MD-2 for the detection of bacterial lipopolysaccharide (LPS) which is an important component of the outer membrane of Gram-negative bacteria.

Biological function summary

This glycoprotein plays a significant role in the innate immune system by recognizing pathogen-associated molecular patterns (PAMPs). It is part of a receptor complex where CD14 acts alongside LPS binding protein (LPS-R) TLR4 and MD-2 to facilitate pro-inflammatory signaling in response to microbial invasion. By binding to LPS CD14 initiates and amplifies the immune response leading to the production of cytokines and the recruitment of other immune cells to the site of infection.

Pathways

CD14 interacts with the toll-like receptor signaling pathway notably involving TLR4. Upon LPS detection CD14 assists in the activation of TLR4 triggering downstream signaling cascades such as NF-κB and MAPK pathways. These pathways result in the transcription of inflammatory cytokines and are essential for the host defense mechanism. The protein complex involving CD14 further interacts with adaptor proteins like MyD88 and TRIF facilitating signal propagation and the immune system's ability to respond to pathogens.

Associated diseases and disorders

Researchers have linked CD14 to sepsis and atherosclerosis. High levels of soluble CD14 (sCD14) are often observed in patients with sepsis indicating its participation in excessive systemic inflammation. In atherosclerosis CD14's role in recognizing LPS contributes to chronic inflammation and the development of plaques in arteries. Through these diseases CD14 connects with TLR4 emphasizing its impact on inflammatory responses and its potential as a therapeutic target.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

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

    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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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), expandable thumbnail

    Western blot - Anti-CD14 antibody [EPR3653] (ab133335)

    Blocking and diluting buffer: 5% NFDM/TBST

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

    All lanes: His-Tagged Human CD14 (aa20 to 345) recombinant protein at 0.015 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 40 kDa

    Observed band size: 43 kDa

    Exposure time: 1s

  • Western blot - Anti-CD14 antibody [EPR3653] (ab133335), expandable thumbnail

    Western blot - Anti-CD14 antibody [EPR3653] (ab133335)

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

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

    Lane 1: Human tonsil lysates prepared in RIPA lysis method at 20 µg

    Lane 2: Human tonsil lysates prepared in 1%SDS Hot lysis method at 20 µg

    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: 45s

  • Western blot - Anti-CD14 antibody [EPR3653] (ab133335), expandable thumbnail

    Western blot - Anti-CD14 antibody [EPR3653] (ab133335)

    Blocking and diluting buffer: 5% NFDM/TBST

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

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

    Lane 1: Human tonsil tissue lysate prepared in 1% SDS Hot lysis method at 20 µg

    Lane 2: HeLa (Human cervix adenocarcinoma) whole cell lysate at 20 µg

    Lane 3: U-937 (Human histiocytic lymphoma) whole cell lysate at 20 µg

    Secondary

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

    Lanes 2 - 3: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 40 kDa

    Observed band size: 53 kDa

    Exposure time: 3min

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

    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), expandable thumbnail

    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), expandable thumbnail

    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), expandable thumbnail

    Western blot - Anti-CD14 antibody [EPR3653] (ab133335)

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

    Lane 1: PBMC cell lysate prepared in 1% SDS Hot lysis method at 10 µg

    Lane 2: Human tonsil tissue lysate prepared in 1%SDS Hot lysis method at 10 µg

    Lane 3: SW480 cell lysate prepared in 1%SDS Hot lysis method at 10 µg

    Secondary

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

    Predicted band size: 40 kDa

    Observed band size: 53 kDa

  • OI-RD Scanning - Anti-CD14 antibody [EPR3653] (ab133335), expandable thumbnail

    OI-RD Scanning - Anti-CD14 antibody [EPR3653] (ab133335)

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

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

    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 (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

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Product protocols

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