リコンビナント
RabMAb

Anti-Thrombomodulin 抗体 [EPR4051] (ab109189)

製品の概要

  • 製品名
    Anti-Thrombomodulin antibody [EPR4051]
    Thrombomodulin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR4051] to Thrombomodulin
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: Flow Cyt, WB, IP, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Synthetic peptide within Human Thrombomodulin (C terminal). The exact sequence is proprietary.

  • ポジティブ・コントロール
    • THP-1, Human placenta and Human heart lysates; Human placenta tissue, Human squamous cervical carcinoma tissue; A431 cells
  • 特記事項

    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.

    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.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
Flow Cyt 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.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    Endothelial cells are unique in synthesizing thrombomodulin.
  • 関連疾患
    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.
  • 配列類似性
    Contains 1 C-type lectin domain.
    Contains 6 EGF-like domains.
  • 翻訳後修飾
    N-glycosylated.
    The iron and 2-oxoglutarate dependent 3-hydroxylation of aspartate and asparagine is (R) stereospecific within EGF domains.
  • 細胞内局在
    Membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • 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.
  • 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.

  • 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 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.

  • Unpurified ab109189, at 1/100 dilution, staining Thombomodulin in A431 cells by Immunofluorescence.

  • 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

  • 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

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

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

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

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

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

  • 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

参考文献

This product has been referenced in:
  • Mazzocchi AR  et al. In vitro patient-derived 3D mesothelioma tumor organoids facilitate patient-centric therapeutic screening. Sci Rep 8:2886 (2018). Read more (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 . Read more (PubMed: 29429903) »
See all 6 Publications for this product

レビューと Q&A

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

Abcam user community

Verified customer

投稿 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
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: 21°C

Abcam user community

Verified customer

投稿 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

Mr. Tyler Crowe

Verified customer

投稿 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
No
Specification
Lymph node
Blocking step
Casein as blocking agent for 15 minute(s) · Concentration: 0.25% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

投稿 Aug 03 2015

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.

Read More

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

登録