Overview

  • Product name

  • Description

    Rabbit polyclonal to Thrombin
  • Host species

    Rabbit
  • Specificity

    The antibody will recognize prothrombin and thrombin
  • Tested applications

    Suitable for: ICC/IF, IHC-P, RIA, IP, WB, ELISAmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Full length native protein: Human thrombin purified from human plasma

  • Positive control

    • IHC-P: Human liver FFPE tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab48626 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
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
RIA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.

Target

  • Function

    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.
  • Tissue specificity

    Expressed by the liver and secreted in plasma.
  • Involvement in disease

    Factor II deficiency
    Ischemic stroke
    Thrombophilia due to thrombin defect
    Pregnancy loss, recurrent, 2
  • Sequence similarities

    Belongs to the peptidase S1 family.
    Contains 1 Gla (gamma-carboxy-glutamate) domain.
    Contains 2 kringle domains.
    Contains 1 peptidase S1 domain.
  • Post-translational
    modifications

    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).
  • Cellular localization

    Secreted, extracellular space.
  • Information by UniProt
  • Database links

  • Alternative names

    • 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

Images

  • 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 ab48626, 1µg/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.

  • ICC/IF image of ab48626 stained HepG2 cells. The cells were 4% formaldehyde (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab48626, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

ab48626 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Answer

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results.

The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.

Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab48626. I would also appreciate if you can confirm some further details:

- Could you confirm when the product was ordered? This will help me trace the origin and internal lot details.
- Reviewing the Swiss-prot entry, I would expect a band at ˜65-70 kDa to be observed, however you state that you would expect a band of 30 kDa. Why would you expect a band size of 30 kDa? The antibody should detect thrombin and prothrombin.
- The antibody was generated against full length native protein so it is unlikely to recognize antihrombin.
- Is it possible to obtain an image so I can understand the problem in more detail?
- Was the sample prepared and resolved under reducing and denaturing conditions? The antibody has been characterized under such conditions.

In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

I hope this information is helpful, and I thank you for your cooperation.

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Answer

Thank you for contacting me again. Detecting thrombin specifically is a very hard issue. As thrombin is part of pro thrombin and the proform undergoes activation during blood coagulation therefore it is difficult to differentiate both proteins. We have several products in our catalogue targeting thrombin, but they are all very likely to recognise pro thrombin as well. In order to obtain clearer thrombin bands, using protein degradation inhibitors are highly recommended (to avoid protein degradation products normally showing as lower target bands), as well as anti coagulants. Thrombin is a very special product, with different sites of glycosilation, producing bands at higher molecular weights. So in this particular case, sample preparation is crucial. However, I would be very happy to offer you a credit note for the price of this antibody, valid for a future purchase. I would really like to help you get the better results from the antibody, so please, let us know how you wish to proceed. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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