Name: Anti-CD14 antibody [EPR3653]
Brand: RabMAb
SKU: ab133335
Rating: 4 (1 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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

Alexa Fluor® 594 Carrier Free HRP

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.

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 (K_D)

K_D = 4.70 x 10 ^-12 M
Learn more about K_D
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

Alternative Versions
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)

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.

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

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)

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)

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)

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)

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)

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)

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)

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)

Equilibrium disassociation constant (K_D)
Learn more about K_D

Click here to learn more about K_D
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)

Western blot protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Protocol Booklet

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

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

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

View all Publications for this product

Customer reviews and Q&As

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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

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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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Dissociation constant (KD)

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (22)

Customer reviews and Q&As

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

Dissociation constant (K_D)

Post-translational
modifications