Recombinant 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
Related conjugates and formulations
Overview
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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.
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Positive control
- IHC-P: human colon, placenta and tonsil tissue; WB: Human tonsil tissue lysate, SW480 and PBMCl lysates.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR3653 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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 |
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WB |
1/1000 - 1/5000. Detects a band of approximately 53 kDa (predicted molecular weight: 40 kDa).
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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 |
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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. |
Target
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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
modificationsN- and O- glycosylated. O-glycosylated with a core 1 or possibly core 8 glycan. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 929 Human
- Omim: 158120 Human
- SwissProt: P08571 Human
- Unigene: 163867 Human
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Alternative names
- CD 14 antibody
- CD_antigen=CD14 antibody
- CD14 antibody
see all
Images
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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.
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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 secondBlocking and diluting buffer: 5% NFDM/TBST
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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 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
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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 minutesBlocking and diluting buffer: 5% NFDM/TBST
The expression level in HeLa and U-937 are low (PMID: 9886426 and 15730927)
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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.
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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.
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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.
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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? -
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (22)
ab133335 has been referenced in 22 publications.
- Futami Y et al. Identification of CD14 and lipopolysaccharide-binding protein as novel biomarkers for sarcoidosis using proteomics of serum extracellular vesicles. Int Immunol 34:327-340 (2022). PubMed: 35294531
- Jarosch S et al. ChipCytometry for multiplexed detection of protein and mRNA markers on human FFPE tissue samples. STAR Protoc 3:101374 (2022). PubMed: 35586313
- Lopez DC et al. Multi-spectral immunofluorescence evaluation of the myeloid, T cell, and natural killer cell tumor immune microenvironment in chordoma may guide immunotherapeutic strategies. Front Oncol 12:1012058 (2022). PubMed: 36338744
- Jarosch S et al. Multiplexed imaging and automated signal quantification in formalin-fixed paraffin-embedded tissues by ChipCytometry. Cell Rep Methods 1:100104 (2021). PubMed: 35475000
- Torchio M et al. Case Report: Exceptional Response to Avelumab After Failure of Electrochemotherapy in a Patient With Rapidly Progressive, PD-L1-Negative Merkel Cell Carcinoma. Front Oncol 11:628324 (2021). PubMed: 34221958