Name: Anti-Thrombomodulin antibody [EPR4051]
Brand: RabMAb
SKU: ab109189
Rating: 4 (4 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR4051] to Thrombomodulin
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 647 APC Carrier Free HRP PE

Product name

Anti-Thrombomodulin antibody [EPR4051]
See all Thrombomodulin primary antibodies
Description

Rabbit monoclonal [EPR4051] to Thrombomodulin
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details
Species reactivity

Reacts with: Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- THP-1, Human placenta and Human heart lysates; Human placenta tissue, Human squamous cervical carcinoma tissue; A431 cells
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. Stable for 12 months at -20°C.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR4051
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)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Thrombomodulin protein (ab98989)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109189 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
Flow Cyt (Intra)		Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB	(1)	1/1000 - 1/10000. Predicted molecular weight: 60 kDa.
IP		1/10 - 1/100.
IHC-P	(2)	1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
ICC/IF		1/100 - 1/300.

Notes
Flow Cyt (Intra) Use at an assay dependent concentration. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB 1/1000 - 1/10000. Predicted molecular weight: 60 kDa.
IP 1/10 - 1/100.
IHC-P 1/100 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
ICC/IF 1/100 - 1/300.

Function

Thrombomodulin is a specific endothelial cell receptor that forms a 1:1 stoichiometric complex with thrombin. This complex is responsible for the conversion of protein C to the activated protein C (protein Ca). Once evolved, protein Ca scissions the activated cofactors of the coagulation mechanism, factor Va and factor VIIIa, and thereby reduces the amount of thrombin generated.
Tissue specificity

Endothelial cells are unique in synthesizing thrombomodulin.
Involvement in disease

Defects in THBD are the cause of thrombophilia due to thrombomodulin defect (THR-THBD) [MIM:188040]. A hemostatic disorder characterized by a tendency to thrombosis.
Defects in THBD are a cause of susceptibility to hemolytic uremic syndrome atypical type 6 (AHUS6) [MIM:612926]. An atypical form of hemolytic uremic syndrome. It is a complex genetic disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, renal failure and absence of episodes of enterocolitis and diarrhea. In contrast to typical hemolytic uremic syndrome, atypical forms have a poorer prognosis, with higher death rates and frequent progression to end-stage renal disease. Note=Susceptibility to the development of atypical hemolytic uremic syndrome can be conferred by mutations in various components of or regulatory factors in the complement cascade system. Other genes may play a role in modifying the phenotype.
Sequence similarities

Contains 1 C-type lectin domain.
Contains 6 EGF-like domains.
Post-translational
modifications

N-glycosylated.
The iron and 2-oxoglutarate dependent 3-hydroxylation of aspartate and asparagine is (R) stereospecific within EGF domains.
Cellular localization

Membrane.
Target information above from: UniProt accession P07204 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 7056 Human
- Omim: 188040 Human
- SwissProt: P07204 Human
- Unigene: 2030 Human
Alternative names
- AHUS 6 antibody
- AHUS6 antibody
- BDCA 3 antibody
- BDCA3 antibody
- CD 141 antibody
- CD141 antibody
- CD141 antigen antibody
- Fetomodulin antibody
- Thbd antibody
- THPH12 antibody
- THRM antibody
- Thrombomodulin antibody
- TM antibody
- TRBM_HUMAN antibody
see all

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Immunohistochemical staining of paraffin embedded human lung with purified ab109189 at a working dilution of 1/1000. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Immunocytochemistry/ Immunofluorescence - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Immunofluorescence staining of A431 cells with purified ab109189 at a working dilution of 1/300, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab109189 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Western blot - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Anti-Thrombomodulin antibody [EPR4051] (ab109189) at 1/10000 dilution (purified) + human placenta lysate at 10 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 60 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Flow Cytometry (Intracellular) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling Thrombomodulin with purified ab109189 at 1/150 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
Immunocytochemistry/ Immunofluorescence - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Unpurified ab109189, at 1/100 dilution, staining Thombomodulin in A431 cells by Immunofluorescence.
Immunoprecipitation - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

ab109189 (purified) at 1/90 immunoprecipitating thrombomodulin in 10 μg human placenta whole cell lysate (Lanes 1 and 2, observed at 100 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
Western blot - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

All lanes : Anti-Thrombomodulin antibody [EPR4051] (ab109189) at 1/1000 dilution (unpurified)

Lane 1 : THP-1 cell lysate
Lane 2 : Human placenta lysate
Lane 3 : Human heart lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 60 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Unpurified ab109189, at 1/100 dilution, staining Thrombomodulin in Human placenta tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Unpurified ab109189, at 1/100 dilution, staining Thrombomodulin in Human squamous cervical carcinoma tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Immunohistochemical analysis of paraffin embedded normal Human spleen tissue using unpurified ab109189 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Immunohistochemical analysis of paraffin embedded normal Human tonsil tissue using unpurified ab109189 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Immunohistochemical analysis of paraffin embedded normal Human lung tissue using unpurified ab109189 showing +ve staining.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR4051] (ab109189)This image is courtesy of an anonymous Abreview

Unpurified ab109189 staining Thrombomodulin in Human artery tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 20% serum for 60 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/200) for 16 hours at 4°C. A Biotin-conjugated Goatanti-rabbit polyclonal (1/200) was used as the secondary antibody.
See Abreview
Anti-Thrombomodulin antibody [EPR4051] (ab109189)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab109189? Please let us know so that we can cite the reference in this datasheet.

ab109189 has been referenced in 8 publications.

Somri-Gannam L et al. IGF1R Axis Inhibition Restores Dendritic Cell Antitumor Response in Ovarian Cancer. Transl Oncol 13:100790 (2020). PubMed: 32428851
Bosisio FM et al. Functional heterogeneity of lymphocytic patterns in primary melanoma dissected through single-cell multiplexing. Elife 9:N/A (2020). PubMed: 32057296
Mazzocchi AR et al. In vitro patient-derived 3D mesothelioma tumor organoids facilitate patient-centric therapeutic screening. Sci Rep 8:2886 (2018). PubMed: 29440675
Wang Y et al. Increased infiltration of CD11 c+/CD123+ dendritic cell subsets and upregulation of TLR/IFN-a signaling participate in pathogenesis of oral lichen planus. Oral Surg Oral Med Oral Pathol Oral Radiol 125:459-467.e2 (2018). IHC-P . PubMed: 29429903
Canavan M et al. Enriched Cd141+ DCs in the joint are transcriptionally distinct, activated, and contribute to joint pathogenesis. JCI Insight 3:N/A (2018). PubMed: 30518680
Teasdale JE et al. Cigarette smoke extract profoundly suppresses TNFa-mediated proinflammatory gene expression through upregulation of ATF3 in human coronary artery endothelial cells. Sci Rep 7:39945 (2017). PubMed: 28059114
Bolognesi MM et al. Multiplex Staining by Sequential Immunostaining and Antibody Removal on Routine Tissue Sections. J Histochem Cytochem 65:431-444 (2017). PubMed: 28692376
Navarro S et al. Association of the Thrombomodulin Gene c.1418C>T Polymorphism With Thrombomodulin Levels and With Venous Thrombosis Risk. Arterioscler Thromb Vasc Biol N/A:N/A (2013). WB ; Human . PubMed: 23520161

Customer reviews and Q&As

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1-5 of 5 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (endothelial cells)

Loading amount

10000 cells

Specification

endothelial cells

Gel Running Conditions

Reduced Denaturing

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

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.

Abcam user community

Verified customer

Submitted Mar 11 2013

Application

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

Sample

Human Tissue sections (artery)

Specification

artery

Fixative

Formaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: citrate

Permeabilization

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 21°C

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.

Abcam user community

Verified customer

Submitted Feb 28 2013

Application

Immunohistochemistry (Frozen sections)

Sample

Human Tissue sections (Human skin)

Permeabilization

Yes - 0.5% Triton TBS Ca/Azide

Specification

Human skin

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

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. Tyler Crowe

Verified customer

Submitted Jul 16 2018

Application

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

Sample

Human Tissue sections (Lymph node)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris-EDTA

Permeabilization

Specification

Lymph node

Blocking step

Casein as blocking agent for 15 minute(s) · Concentration: 0.25% · Temperature: RT°C

Fixative

Formaldehyde

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.

Abcam user community

Verified customer

Submitted Aug 03 2015

Question

Por favor, es urgente saber si el anticuerpo EPCR4051, ab109189 (anti-trombomodulina) puede usarse para western blotting en condiciones reductoras, o solamente sin desnaturalizar.Muchas gracias.

Abcam community

Verified customer

Asked on Jan 07 2013

Answer

Gracias por contactarnos.

Salvo que se especifique lo contrario en la ficha técnica del anticuerpo, todos nuestros anticuerpos reconocen el epítopo bajo condiciones desnaturalizantes y reductoras.

Para ello, se recomienda usar buffer RIPA para lisar las células, por su alto poder desnaturalizante (ver receta https://www.abcam.com/index.html?pageconfig=resource&rid=11379) y usar un agente reductor en el buffer de carga (ß-mercaptoethanol o DTT) calentando las muestras previamente a 95-100C durante 5-10 minutos.

Si quiere más información sobre la técnica, nuestro https://www.abcam.com/index.html?pageconfig=resource&rid=11375 puede resultar útil.

Espero que esta información sea de utilidad. Para cualquier otra consulta no dude en contactarnos de nuevo.

Le recuerdo que todos nuestros productos tienen 6 meses de garantía, y en caso de o comportarse como se especifica en la ficha técnica, se reemplazará o reembolsará el importe del mismo.

Abcam Scientific Support

Answered on Jan 07 2013

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

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (8)

Customer reviews and Q&As

Western blot abreview for Anti-Thrombomodulin antibody [EPR4051]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Thrombomodulin antibody [EPR4051]

Immunohistochemistry (Frozen sections) abreview for Anti-Thrombomodulin antibody [EPR4051]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Thrombomodulin antibody [EPR4051]

Post-translational
modifications