Anti-Thrombin 抗体 [5G9] (ab17199)

製品の概要

  • 製品名Anti-Thrombin antibody [5G9]
    Thrombin 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [5G9] to Thrombin
  • 特異性Binds to human thrombin and to a complex of human thrombin with the thrombin inhibitor hirudin.
  • アプリケーション適用あり: ELISA, WB, IHC-Pmore details
  • 種交差性
    交差種: Cow, Human
  • 免疫原

    Full length native protein (purified) corresponding to Human Thrombin. Thrombin isolated from activated human plasma

  • エピトープnot mapped
  • ポジティブ・コントロール
    • purified alpha thrombin IHC-P: Human liver FFPE tissue sections.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab17199 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
ELISA 1/800.
WB 1/500. Detects a band of approximately 75 kDa (predicted molecular weight: 70 kDa).
IHC-P Use a concentration of 10 µg/ml.

ターゲット情報

  • 機能Thrombin, which cleaves bonds after Arg and Lys, converts fibrinogen to fibrin and activates factors V, VII, VIII, XIII, and, in complex with thrombomodulin, protein C. Functions in blood homeostasis, inflammation and wound healing.
  • 組織特異性Expressed by the liver and secreted in plasma.
  • 関連疾患Factor II deficiency
    Ischemic stroke
    Thrombophilia due to thrombin defect
    Pregnancy loss, recurrent, 2
  • 配列類似性Belongs to the peptidase S1 family.
    Contains 1 Gla (gamma-carboxy-glutamate) domain.
    Contains 2 kringle domains.
    Contains 1 peptidase S1 domain.
  • 翻訳後修飾The gamma-carboxyglutamyl residues, which bind calcium ions, result from the carboxylation of glutamyl residues by a microsomal enzyme, the vitamin K-dependent carboxylase. The modified residues are necessary for the calcium-dependent interaction with a negatively charged phospholipid surface, which is essential for the conversion of prothrombin to thrombin.
    N-glycosylated. N-glycan heterogeneity at Asn-121: Hex3HexNAc3 (minor), Hex4HexNAc3 (minor) and Hex5HexNAc4 (major). At Asn-143: Hex4HexNAc3 (minor) and Hex5HexNAc4 (major).
  • 細胞内局在Secreted, extracellular space.
  • Information by UniProt
  • 参照データベース
  • 別名
    • Coagulation factor II antibody
    • Coagulation factor II thrombin antibody
    • EC 3.4.21.5 antibody
    • F2 antibody
    • Factor II antibody
    • Prepro coagulation factor II antibody
    • Prothrombin antibody
    • Prothrombin B chain antibody
    • PT antibody
    • RPRGL2 antibody
    • Serine protease antibody
    • THPH1 antibody
    • THRB_HUMAN antibody
    • Thrombin heavy chain antibody
    see all

Anti-Thrombin antibody [5G9] 画像

  • ab17199 staining Thrombin in human liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde, permeabilized with 0.2% Triton X-100 in PBS and blocked with 5% milk for 30 minutes at room temperature; antigen retrieval was by heat mediation in Tris pH 9.0. Samples were incubated with primary antibody (1/100 in PBS) for 16 hours at 4°C. An undiluted Biotin-conjugated horse anti-mouse IgG polyclonal was used as the secondary antibody.

    See Abreview

  • IHC image of Thrombin staining in human liver formalin fixed paraffin embedded tissue section, performed on a Leica Bond system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab17199, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Thrombin antibody [5G9] (ab17199) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Human Plasma Total Protein Lysate
    Lane 3 : HepG2 nuclear extract lysate (ab14660)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 75 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 40 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
    Thrombin contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

Anti-Thrombin antibody [5G9] (ab17199) 使用論文

ab17199 has not yet been referenced specifically in any publications.

Product Wall

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: HIER Tris/HCL pH 9.0
Sample Cow Tissue sections (Bovine Liver)
Specification Bovine Liver
Permeabilization Yes - 0.2% Triton X-100 and PBS
Fixative Formaldehyde
Username

Abcam user community

Verified customer

投稿 Aug 08 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris pH 9.0
Sample Human Tissue sections (human liver)
Specification human liver
Permeabilization Yes - 0.2% Triton X-100 and PBS
Fixative Formaldehyde
Username

Dr. Steffen Rickelt

Verified customer

投稿 Mar 19 2014

Please find the CoC forab17199 attached as requested.

If you have any further questions please do not hesitate to ask.

Thank you for contacting us.

Please find attached the new CoC as requested.

If you have any further requests, please do not hesitate to contact us.

A western blot which is unfortunately unavailable shows ab17199 can detect a 36kDa band and a 31 and a6kDa under reducing conditions. I have looked at western blots from other thrombinantibodies on our catalog and they also appear to detect bands aroun...

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Thank you for your enquiry.

I have checked the SwissProt entry for humanthrombin at http://www.uniprot.org/uniprot/P00734.

This link contains a thrombin sequence of 622 amino acids with predicted molecular weight of about 70kDa. Thi...

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Thank you for contacting us.
This antibody recognizes Thrombin and not ProThrombin. The higher than expected molecular weight that is seen in western blotting could be due to the glycosylation that Thrombin undergoes (based on the sequence form Sw...

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Thank you for contacting Abcam.

I have talked to the supplying lab and they have confirmed that this antibody only recognizes Thrombin and not ProThrombin. The higher than expected molecular weight that is seen in western blotting could be du...

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Thank you for your inquiry and I apologize for the delay in this reply. Yes, ab18447 is a suitable isotype control for ab17199. http://www.abcam.com/Mouse-IgG1-kappa-monoclonal-MG1-45-Isotype-control-ab18447.html I hope this inform...

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Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. Thrombin following processing from its precursor prothrombin has a molecular weight of 36KDa (please see the wikipedia link below). However, t...

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