Anti-Thrombomodulin antibody [PBS-01] (ab6980)

Reviews (7)Q&A (9)References (17)

Overview

  • Product name

    Anti-Thrombomodulin antibody [PBS-01]
    See all Thrombomodulin primary antibodies

  • Description

    Mouse monoclonal [PBS-01] to Thrombomodulin

  • Host species

    Mouse

  • Tested applications

    Suitable for: ELISA, WB, Flow Cyt, IHC-Pmore details

  • Species reactivity

    Reacts with: Rabbit, Human
    Does not react with: Pig

  • Immunogen

    Recombinant full length protein (Human).

  • Epitope

    EGF domain 5 of thrombomodulin.

Properties

Applications

Our Abpromise guarantee covers the use of ab6980 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
ELISA Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • 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.
  • Information by UniProt

  • Database links

    • 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

    Images

    • Western blot - Anti-Thrombomodulin antibody [PBS-01] (ab6980)
      Western blot - Anti-Thrombomodulin antibody [PBS-01] (ab6980)Image courtesy of an anonymous Abreview.
      Anti-Thrombomodulin antibody [PBS-01] (ab6980) at 1/100 dilution + whole cell lysate prepared from human aortic endothelial cells at 30 µg

      Secondary
      HRP conjugated anti-mouse IgG at 1/4000 dilution

      Developed using the ECL technique.

      Observed band size: 95 kDa
      why is the actual band size different from the predicted?


      Exposure time: 10 minutes


      36kDa band is GAPDH in the same lane.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [PBS-01] (ab6980)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Thrombomodulin antibody [PBS-01] (ab6980)
      Ab6980 staining human normal lung endothelium. Staining is localized to the membrane.
      Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
      Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
    • Flow Cytometry - Anti-Thrombomodulin antibody [PBS-01] (ab6980)
      Flow Cytometry - Anti-Thrombomodulin antibody [PBS-01] (ab6980)
      Overlay histogram showing THP1 cells stained with ab6980 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab6980, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in THP1 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    References

    This product has been referenced in:

    • Suzuki K  et al. Neutrophil Elastase Damages the Pulmonary Endothelial Glycocalyx in Lipopolysaccharide-Induced Experimental Endotoxemia. Am J Pathol 189:1526-1535 (2019). Read more (PubMed: 31108101) »
    • Sobol N  et al. Synthesis and targeting of gold-coated 177Lu-containing lanthanide phosphate nanoparticles-A potential theranostic agent for pulmonary metastatic disease. APL Bioeng 2:016101 (2018). Read more (PubMed: 31069286) »
    See all 17 Publications for this product

    Publishing research using ab6980? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 16 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Thrombomodulin antibody [PBS-01]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (bladder)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH9.0
    Permeabilization
    No
    Specification
    bladder
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 27 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Thrombomodulin antibody [PBS-01]

     Poor
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Baboon Tissue sections (Heart)
    Antigen retrieval step
    Enzymatic - Buffer/Enzyme Used: Proteinase K
    Permeabilization
    No
    Specification
    Heart
    Blocking step
    Serum Free Protein Block (Dako) as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C
    Fixative
    10% NBF
    Read More

    Premier Laboratory

    Verified customer

    Submitted Apr 26 2016

    Immunohistochemistry (Frozen sections) abreview for Anti-Thrombomodulin antibody [PBS-01]

     Inconclusive
    Abreviews
    This product is known to not work in this application or species.
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Pig Tissue sections (Heart)
    Permeabilization
    No
    Specification
    Heart
    Blocking step
    Serum Free Protein Block (Dako) as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C
    Fixative
    Acetone
    Read More

    Premier Laboratory

    Verified customer

    Submitted Apr 19 2016

    Immunohistochemistry (Frozen sections) abreview for Anti-Thrombomodulin antibody [PBS-01]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (heart and lung)
    Permeabilization
    No
    Specification
    heart and lung
    Blocking step
    Serum Free Protein Block (Dako) as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C
    Fixative
    Acetone
    Read More

    Premier Laboratory

    Verified customer

    Submitted Apr 19 2016

    Question

    I just have a quick question. What part of thrombomodulin does this antibody (ab6980) bind to? (i.e. C or N-terminal)
    Thank you,

    Read More
    Answer

    Thank you for contacting us.

    ab6980 binds within the EGF 5-6 domain of human TM, rather distant from the N-terminus of the protein.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    The project I am working on involves using your Anti-thrombomodulin antibody (ab6980) for immunohistochemistry of canine samples. Your data sheet states this antibody can be used for canine samples, but the advisory committee is asking for proof/references that it is actually detecting the thrombomodulin and not something else in canine samples. Do you have any references that can help me answer this question, or can you suggest how to find this information?

    Read More
    Answer

    Thank you for your patience as I have looked into your inquiry.


    Unfortunately, I have not been able to obtain any additional information from the originating laboratory regarding references or additional data in which this antibody was used in canine samples. I do apologize for any inconvenience this may cause.


    Please do not hesitate to contact me if you have any additional questions.

    Read More

    Question

    sorry about the confusion, I'm not sure which number did you mean by original abcam order number so I gave you the purchase order number from the pack slip.

    So here it is, the purchase order number is *****, and there is an order number also, which is: ***, the customer is ******, the shipping date is ******.
    also attached a picture of the packing slip from abcam if that's helpful. Again I'm sorry for the confusion and if you need any other information, please let me know. Thank you so much!

    Read More
    Answer

    Thank you for your reply and apologies for the confusion when I asked for the original Abcam order information, I meant the order information for ab6980.

    The second order number you provided was exactly what I needed and I have now sent you ab94373 as a free of charge replacement. Your new order number is ******.

    Please let me know if there is anything else I can help you with.

    Read More

    Question

    I ran the blot as you suggested using 5% BSA as blocking reagent and loaded 50 µg of protein but the blot still came out to have multiple bands like before.
    It would be really nice if you could send us an replacement antibody and I'd really appreciate it.

    The original order number for this mouse monoclonal thrombomodulin antibody is: ******, customer ******.

    And the catalogue number of the antibody I'd like as replacement is ab94373, it's a rabbit polyclonal antibody.

    Thank you so much for your help!

    Read More
    Answer

    Thank you for your reply.

    As mentioned previously we do guarantee our products for 6 months from the date of delivery, I mention this as the order number you provided is from September 2012 and ab6980 was not on that order (also the customer name you provided does not match the information that is on that order number you gave).

    Could you please check through your records to find the correct order number and also could you please let me know which institution you are working for.

    I look forward to your reply.

    Read More

    Question

    Hi, there, thank you for your prompt response.
    To answer your questions:
    1. I see multiple bands ranging from molecular weight 20 KDa all the way to 120 KDa. They are distinct bands but these bands will only show up if I use the super sensitive substrate to do the detection.
    2. The most protein I've loaded was 50 µg and that's the sample that gave me the multiple bands. The lanes that I loaded 10 µg, 20 µg of protein didn't have any bands on them.
    3. I use 5% non-fat dry milk in TBST as blocking agent.
    4. I've tried 1:1000, 1:500, and even 1:200 dilutions and I do overnight incubation at 4℃ for the primary antibody incubation.

    Thank you so much! Have a great weekend!

    Read More
    Answer

    Thank you for your reply.

    As mentioned previously, this antibody is covered under our Abpromise and so if we cannot help resolve these issues, then ill be happy to either replace or refund this antibody.

    Based on the information you have provided, loading 50ug of lysate and switching to a 5%BSA block, to help prevent non-specific binding may prove to be successful.

    If this does not work or if you feel it may not work, then please let me know and also let me know and provide the original Abcam order number and the catalogue number of the antibody you would like as a replacement and I will be happy to send it out.

    If there is anything else I can help you with, just let me know.

    Read More

    Question

    Hi, I'm writing to you about the anti-thrombomodulin mouse monoclonal antibody we purchased from abcam. I used this antibody on HUVECs cell lysate to blot for thrombomodulin and it hasn't worked for me. I tried different dilutions of the antibody( up to 1:200) and loaded different amount of cell lysate and there were no bands at the 94 molecular weight... and I tried to use a super sensitive substrate to do the exposure, and there are a smear of multiple bands on the blot...
    I'm just wondering what would you advice to use this antibody for western blot and if it's possible to get an replacement for it.
    Thank you so much for your help!

    Read More
    Answer

    Thank you for contacting Abcam.

    The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on human samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

    To see if there is any protocol tips I can suggest that will help resolve the problems, would you be able to answer a few questions:

    1- Are you seeing any bands, I know you said there was a smear, but do you see any distinct bands?

    2- How much sample are you loading?

    3- What type of blocking agent are you using?

    4- What primary antibody concentrations have you tried and how long do you incubate for?

    Looking forward to your reply and helping resolve this issue.

    Hope you have a great weekend.

    Read More

    1-10 of 16 Abreviews or Q&A

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