Recombinant Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

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Western blot - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18217-209] to Thrombomodulin
Suitable for: IHC-Fr, ICC/IF, IP, Flow Cyt, IHC-P, WB
Reacts with: Mouse

Carrier Free

Product name

Anti-Thrombomodulin antibody [EPR18217-209]
See all Thrombomodulin primary antibodies
Description

Rabbit monoclonal [EPR18217-209] to Thrombomodulin
Host species

Rabbit
Tested applications

Suitable for: IHC-Fr, ICC/IF, IP, Flow Cyt, IHC-P, WBmore details
Species reactivity

Reacts with: Mouse
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Mouse lung and placenta tissue lysates; bEND.3 whole cell lysate. IHC-P: Mouse lung and stomach tissues. IHC-Fr: Mouse embryo E14.5 (developing lung) tissue. ICC/IF: bEND.3 cells. Flow Cyt: bEND.3 cells. IP: bEND.3 whole cell lysate; Mouse lung tissue lysate.
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.

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.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR18217-209
Isotype

IgG
Research areas

Alternative Versions
- Anti-Thrombomodulin antibody [EPR18217-209] - BSA and Azide free (ab230152)
Compatible Secondaries
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)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab230010 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
IHC-Fr		1/500. Perform heat mediated antigen retrieval by using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF		1/100.
IP		1/30.
Flow Cyt		1/500.
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/1000. Detects a band of approximately 75,105 kDa (predicted molecular weight: 62 kDa).

Notes
IHC-Fr 1/500. Perform heat mediated antigen retrieval by using sodium citrate buffer (10 mM citrate pH 6.0 + 0.05% Tween-20).
ICC/IF 1/100.
IP 1/30.
Flow Cyt 1/500.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 75,105 kDa (predicted molecular weight: 62 kDa).

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: 21824 Mouse
- SwissProt: P15306 Mouse
- Unigene: 24096 Mouse
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

Western blot - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

All lanes : Anti-Thrombomodulin antibody [EPR18217-209] (ab230010) at 1/1000 dilution

Lane 1 : Mouse lung tissue lysate at 20 µg
Lane 2 : bEND.3 (mouse brain endothelioma cell line) whole cell lysate at 10 µg
Lane 3 : Mouse placenta tissue lysate at 10 µg

Secondary
Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 2-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Developed using the ECL technique.

Predicted band size: 62 kDa
Observed band size: 105,75 kDa why is the actual band size different from the predicted?

Exposure times: Lane 1: 1 minute; Lane 2: 20 seconds; Lane 3: 10 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.

The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.
Immunocytochemistry/ Immunofluorescence - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized bEND.3 (mouse brain endothelioma cell line) cells labeling Thrombomodulin with ab230010 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and membranous staining in bEND.3 cell line (PMID: 7622601; PMID: 8223719).

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Immunoprecipitation - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

Thrombomodulin was immunoprecipitated from 0.35 mg of mouse lung tissue lysate with ab230010 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230010 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: Mouse lung tissue lysate 10 μg (Input).
Lane 2: ab230010 IP in mouse lung tissue lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230010 in mouse lung tissue lysate.

Exposure time: 10 seconds.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Immunoprecipitation - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

Thrombomodulin was immunoprecipitated from 0.35 mg of bEND.3 (mouse brain endothelioma cell line) whole cell lysate with ab230010 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab230010 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: bEND.3 whole cell lysate 10 μg (Input).
Lane 2: ab230010 IP in bEND.3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab230010 in bEND.3 whole cell lysate.

Exposure time: 10 seconds.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Flow Cytometry - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

Flow cytometric analysis of bEND.3 (mouse brain endothelioma cell line) cells labeling Thrombomodulin with ab230010 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

Gated on total viable cells.
Immunohistochemistry (Frozen sections) - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse embryo E14.5 (developing lung) tissue labeling Thrombomodulin with ab230010 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution (green). Positive membrane staining in the developing lung in mouse E14.5 embryo (PMID: 28306049) is observed.

The nuclear counterstain is DAPI (blue).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling Thrombomodulin with ab230010 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on endothelial cells of mouse lung (PMID: 23946288; PMID: 10231031) is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Thrombomodulin with ab230010 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on endothelial cells of mouse stomach (PMID: 23946288; PMID: 10231031) is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-Thrombomodulin antibody [EPR18217-209] (ab230010)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

Download

Publishing research using ab230010? Please let us know so that we can cite the reference in this datasheet.

ab230010 has been referenced in 2 publications.

Kondo T et al. Immunohistochemical analysis of thrombomodulin expression in myocardial tissue from autopsy cases of ischemic heart disease. Leg Med (Tokyo) 51:101897 (2021). PubMed: 33940278
Dobie R et al. Single-Cell Transcriptomics Uncovers Zonation of Function in the Mesenchyme during Liver Fibrosis. Cell Rep 29:1832-1847.e8 (2019). PubMed: 31722201

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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

Related Products

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

Certificate of Compliance

References (2)

Customer reviews and Q&As

Post-translational
modifications