Overview

  • Product name

    Anti-Von Willebrand Factor antibody
    See all Von Willebrand Factor primary antibodies
  • Description

    Sheep polyclonal to Von Willebrand Factor
  • Host species

    Sheep
  • Tested applications

    Suitable for: IHC-Fr, Immunodiffusion, Flow Cyt, ELISA, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Human, Pig
  • Immunogen

    Purified human von Willebrand factor.

  • Positive control

    • Tonsil.
  • General notes

    This product should be stored undiluted. Storage in frost free freezers is not recommended. Should this product contain a precipitate we recommend microcentrifugation before use.

Properties

Applications

Our Abpromise guarantee covers the use of ab11713 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 1/50 - 1/100.
Immunodiffusion 1/1.
Flow Cyt Use at an assay dependent concentration.

ab37385 - Sheep polyclonal IgG, is suitable for use as an isotype control with this antibody.

ELISA Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function

      Important in the maintenance of hemostasis, it promotes adhesion of platelets to the sites of vascular injury by forming a molecular bridge between sub-endothelial collagen matrix and platelet-surface receptor complex GPIb-IX-V. Also acts as a chaperone for coagulation factor VIII, delivering it to the site of injury, stabilizing its heterodimeric structure and protecting it from premature clearance from plasma.
    • Tissue specificity

      Plasma.
    • Involvement in disease

      Defects in VWF are the cause of von Willebrand disease (VWD) [MIM:277480]. VWD defines a group of hemorrhagic disorders in which the von Willebrand factor is either quantitatively or qualitatively abnormal resulting in altered platelet function. Symptoms vary depending on severity and disease type but may include prolonged bleeding time, deficiency of factor VIII and impaired platelet adhesion. Type I von Willebrand disease is the most common form and is characterized by partial quantitative plasmatic deficiency of an otherwise structurally and functionally normal Willebrand factor; type II is associated with a qualitative deficiency and functional anomalies of the Willebrand factor; type III is the most severe form and is characterized by total or near-total absence of Willebrand factor in the plasma and cellular compartments, also leading to a profound deficiency of plasmatic factor VIII.
    • Sequence similarities

      Contains 1 CTCK (C-terminal cystine knot-like) domain.
      Contains 4 TIL (trypsin inhibitory-like) domains.
      Contains 3 VWFA domains.
      Contains 3 VWFC domains.
      Contains 4 VWFD domains.
    • Domain

      The von Willebrand antigen 2 is required for multimerization of vWF and for its targeting to storage granules.
    • Post-translational
      modifications

      All cysteine residues are involved in intrachain or interchain disulfide bonds.
      N- and O-glycosylated.
    • Cellular localization

      Secreted. Secreted > extracellular space > extracellular matrix. Localized to storage granules.
    • Information by UniProt
    • Database links

    • Alternative names

      • Coagulation factor VIII antibody
      • Coagulation factor VIII VWF antibody
      • F8VWF antibody
      • Factor VIII related antigen antibody
      • von Willebrand antigen 2 antibody
      • von Willebrand antigen II antibody
      • Von Willebrand disease antibody
      • Von Willebrand factor precursor antibody
      • VWD antibody
      • vWF antibody
      • VWF_HUMAN antibody
      see all

    Images

    • ab11713 at a 1/1000 dilution staining Von Willebrand Factor in mouse blood vessels by Immunohistochemistry (frozen sections) incubated for 16 hours at 4°C. PFA fixed. Permeabilized with 0.1% Triton. Blocked using 20% serum for 1 hour at 19°C. Secondary used at 1/200 polyclonal Donkey anti-sheep conjugated to Alexa Fluor 647 (pink). Counterstain (blue) DAPI.

      See Abreview

    • ab11713 staining Von Willebrand Factor in porcine heart tissue by Immunohistochemistry (Frozen sections).
      Tissue was fixed in acetone, permeabilized using 0.3% Triton/PBS, blocked with 10% serum for 30 minutes at 22°C and then incubated with ab11713 at a 1/50 dilution for 16 hours at 4°C. The secondary used was a FITC conjugated donkey anti-sheep polyclonal used at a 1/1000 dilution.

      See Abreview

    References

    This product has been referenced in:

    • Patil R  et al. Polymalic acid chlorotoxin nanoconjugate for near-infrared fluorescence guided resection of glioblastoma multiforme. Biomaterials 206:146-159 (2019). Read more (PubMed: 30933776) »
    • Perdomo J  et al. Neutrophil activation and NETosis are the major drivers of thrombosis in heparin-induced thrombocytopenia. Nat Commun 10:1322 (2019). Read more (PubMed: 30899022) »
    See all 45 Publications for this product

    Customer reviews and Q&As

    11-20 of 25 Abreviews or Q&A

    Answer

    Hello! The general protocol for Flow Cytometry is below. The reference for the paper is:
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2195953/
    Please let me know if you have any further questions!
    Cells Flow cytometry intracellular staining protocol
    General procedure describing detection of intracellular proteins in flow cytometry
    Fixing and permeabilization:
    For intracellular staining, cells can be fixed first to ensure stability of soluble antigens or antigens with a short half life (see the special recomendations below for important exceptions). This should retain the target protein in the original cellular location.
    Detection of intracellular antigens requires a cell permeabilization step prior to staining. Antibodies should be prepared in permeabilization buffer to ensure the cells remain permeable. When gating on cell populations, the light scatter profiles of the cells on the flow cytometer will change considerably after permeabilization.
    N/B Cell surface staining should be performed prior to fixation.
    There are several methods available:
    1. Formaldehyde followed by detergent:
    Fixation in 0.01% formaldehyde for 10-15 min (this will stabilize proteins), followed by disruption of membrane by detergent.
    Detergents:
    Triton or NP-40 (0.1 to 1% in PBS). These will also partially dissolve the nuclear membrane and are therefore very suitable for nuclear antigen staining. It should be noted that loss of cell membrane and cytoplasm will result in decreased light scattering and also in reduced non specific fluorescence.
    Tween 20, Saponin, Digitonin and Leucoperm are mild membrane solubilisers. Use at 0.5% v/v in PBS. These give large enough pores for antibodies to go through without dissolving plasma membrane. Suitable for antigens in the cytoplasm or the cytoplasmic face of the plasma membrane. Also suitable for soluble nuclear antigens.
    2. Formaldehyde (0.01%) followed by methanol (SEE 3)
    3. Methanol followed by detergent.
    Add 1 ml ice cold methanol to each sample.
    Mix gently. Place at -20oC for 10 minutes.
    Centrifuge, wash twice in PBS 1% BSA.
    4. Acetone fixation and permeabilization:
    Add 1 ml ice cold acetone to each sample.
    Mix gently. Place at -20oC for t5 to 10 minutes.
    Centrifuge, wash twice in PBS 1% BSA.
    N/B Polystyrene/plastic tubes are not suitable for use with acetone.
    SPECIAL RECOMMENDATIONS:
    Antigens close to the plasma membrane and soluble cytoplasmic antigens will require mild cell permeabilization without fixation.
    Cytoskeletal, viral and some enzyme antigens usually give optimal results when fixed with acetone, alcohol or formaldehyde (high concentration).
    Antigens in cytomplasmic organelles and granules will require a fixation and permeabilization method depending on the antigen. The epitope needs to remain accessible.
    Intracellular staining procedure:
    1.Add 100 µl of fixative. Incubate for 10 minutes at required temperature (see above).
    2.Add 100µl detergent based permeabilizing agent and incubate in the dark at room at room temperature for 15 minutes.
    3.Wash the cells by adding 2ml of PBS (containing 0.1% triton or other permeabilizing detergent), centrifuge at 300g (2000 rpm) for 5 minutes, discard supernatant and re-suspend the pellet in the volume remaining.
    4.Follow antibody staining procedure as indicated in our ‘direct’ and ‘indirect’ protocols.
    Antibodies should be prepared in permeabilization buffer to ensure the cells remain permeable.
    Detection of secreted proteins:
    Detection of secreted proteins is difficult as the protein will be released from the cell before detection, or may degrade rapidly. Brefaldin A and other compounds are often used as a Golgi-Block. Cells are incubated with Brefaldin A which prevents proteins being released from the golgi. Any cells expressing the protein can then be detected.

    Read More
    This product is known to not work in this application or species.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Lung)
    Specification
    Lung
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

    Mr. Carl Hobbs

    Verified customer

    Submitted Sep 07 2012

    Answer

    Thank you for contacting us and your interest in our products.

    We have the information that this antibody has been used to detect the Von Willebrand Factor of cultured lymphatic and blood vessel endothelial cells using flow cytometry. This was performed by isolating the cell, performing fixation and permiabilisation, then incubating them with the antibody. A FITC conjugated secondary antibody was used for detection.

    We have a number of secondary antibodies which may be suitable for your experiments, here are two examples:

    Donkey anti-Sheep IgG H&L (PE) secondary antibodyhttps://www.abcam.com/Sheep-IgG-secondary-antibody-H-L-ab7009.html

    Donkey polyclonal Secondary Antibody to Sheep IgG - H&L (DyLight® 550) https://www.abcam.com/Donkey-polyclonal-Secondary-Antibody-to-Sheep-IgG-H-L-DyLight-550-ab96940.html

    More can be found from the following link:

    https://www.abcam.com/index.html?pageconfig=searchresults&search=igg&pt=7&sd=4&fViewMore=1

    The following isotype control should be suitable:

    Sheep IgG - Isotype Control (https://www.abcam.com/Sheep-IgG-Isotype-Control-ab37385.html).

    I hope this information has been of help. If you have any further questions, please do not hesitate to contact us again.

    Read More
    This product is known to not work in this application or species.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Adult Lung)
    Specification
    Adult Lung
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted May 21 2012

    Answer

    Thank you for your inquiry.

    The total protein concentration of ab11713 (lot 1280839) is 9.13mg/ml.

    This product is purified IgG which was prepared from serum by ion exchange chromatography. Due to the nature of ion exchange chromatography, various proteins will be purified out of serum along with IgG. The concentration of specific antibody in this preparation is ˜10% of the total protein.

    I hope this information is helpful and wish you good luck with your experiments.

    Read More

    Answer

    Vielen Dank für Ihren Anruf.

    Wie versprochen habe ich bezüglich des Von Willebrand FactorAntikörpers ab11713 im Labor nachgefragt. Ich kann Ihnen nun bestätigen, dass dieser Antikörper spezifisch für den Von Willebrand Factorist (SwissProt ID P04275,http://www.uniprot.org/uniprot/P04275). Die Angabe von Factor VIII (eine Mischung der alternativen Namen VWF und 'Factor VIII related antigen') war irreführend und wurde vom Datenblatt entfernt.

    Leider bin ich mir nicht ganz sicher, welcher unserer VWF-Antikörper am besten für Ihre Versuchsvorhaben geeignet ist:

    Wir hätten zum Beispiel den Maus-monoklonalen ab68545: https://www.abcam.com/index.html?datasheet=68545 (or use the following: https://www.abcam.com/index.html?datasheet=68545). Allerdings ist das Epitop leider nicht kartiert worden, daher kann ich nicht über eine Interaktion mit Factor VIII (und wie sich das für Ihre Versuche auswirkt) spekulieren.

    Des Weiteren hätten wir den Schaf-polyklonalen ab11713 und den Ziegen-polyklonalen ab115771, die beide gegen (aus Plasma) aufgereinigten VWF generiert wurden, und daher vielleicht für Ihre Plasma-/Serummessungen geeignet sein könnten:
    https://www.abcam.com/index.html?datasheet=11713 (or use the following: https://www.abcam.com/index.html?datasheet=11713).
    https://www.abcam.com/index.html?datasheet=115771 (or use the following: https://www.abcam.com/index.html?datasheet=115771).

    Ich möchte Ihnen jedoch versichern, dass alle diese Antikörper spezifisch für VWF sind (und nicht mit Factor VIII kreuzreagieren) sowiefür ELISA getestet wurden. Wir geben alle getesteten Anwendungen auf unseren Produktdatenblättern an undgarantieren, dass der Antikörper in allen diesen Anwendungenfunktioniert.Falls sich herausstellt, dass er nicht so funktioniert, wie auf dem Datenblatt beschrieben und Sie sich innerhalb von sechs Monaten mit Ihrem Problem an uns wenden, werden wir Ihnen gerne Vorschläge zur Optimierung Ihres Protokolls oder gegebenenfalls einen Ersatz oder eine Gutschrift schicken (https://www.abcam.com/Abpromise).

    Außerdem können wir Ihnen ein spezielles Angebot über einen 100%igen Abreview-Rabatt machen: Da wir derzeitkeine Abbildung von den oben genannten Antikörpern im ELISA haben, können Sie an unserem Testprogramm teilnehmen.Dabei bekommen Sieeinen kostenlosenPrimärantikörper von uns,wenn Sie uns ein Abreview mit dem Testresultat und einer Abbildung zusenden. Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

    1.) Bestätigen Sie mir bitte, welchen VWF-Antikörper Sie kaufen möchten und dass bereit sind uns eine Abbildung Ihrer Ergebnisse zukommen zu lassen.

    2.) Bitte bestellen Sie erst nach Erhalt des Rabattcodes.

    3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

    4.) Testen Sie den Antikörper im ELISA.

    5.) Senden Sie uns die Abbildung Ihres Ergebnisses mittels eines Abreviews zu und notieren Sie den Discount Code in dem Feld "additional notes". Unter der folgenden URL können Sie mehr über unser Abreview System erfahren: https://www.abcam.com/abreviews

    6.) Der Rabattcode ist nach dem Abschicken des Abreviews aktiv, und Sie können einen anderen primären Antikörper bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode und die Bestellnummer bereit). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

    Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
    https://www.abcam.com/collaborationdiscount

    Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

    Read More

    Question
    Answer

    Thank you for your call today and for your questions about our VWF antibodies.
    As we discussed, ab68545 would be a good antibody to try in sandwich ELISA with ab6994. I have issued a discount code that is worth 1 free primary antibody after submitting an Abreview with the data from this sandwich ELISA, and the details are below. We also discussed a purification kit to use before conjugating HRP to the detection antibody, and ab102783 would be a good kit to use with ab68545-
    https://www.abcam.com/Antibody-Purification-kit-1-purification-ab102783.html
    Please see below for your discount code, and let me know if you have any questions or if there is anything else that I can do for you.
    DISCOUNT CODE: ***
    Expiration date: July 13, 2012
    I am very pleased to hear you would like to test ab68545 in a sandwich ELISA with ab6994. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab68545 in sELISA with ab6994, and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
    The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.
    Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.
    The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

    Read More

    Answer

    Thank you for getting back to me and for gathering more information about the customer's assay.

    I have conducted some search for your customer and found some products which might be suitable for this application. However, it is important to emphasize that none of our antibodies againstvon Willebrand factor have been tested in antibody-coated latex beads. Please advise your customer to take a look at the on-line product datasheets carefully:

    ab68545: https://www.abcam.com/von-willebrand-factor-antibody-2q2134-ab68545.html

    ab11713: https://www.abcam.com/von-willebrand-factor-antibody-ab11713.html

    ab8820: https://www.abcam.com/von-willebrand-factor-antibody-ab8820.html

    ab115771: https://www.abcam.com/von-willebrand-factor-antibody-ab115771.html

    I hope this helps and if I can assist further, please do not hesitate to contact me.

    Read More

    Question
    Answer

    Thank you for contacting us.
    We have many antibodies that specifically stain endothelial and osteoblast marker proteins. The most popular antibodies are as follows;
    Osteocalcin, DMP-1 and Osteopontin are Osteoblast marker proteins, the antibodies against these are;
    ab13420, ab13418, ab13421, ab14173
    ab103203, ab81985, ab76632 and 82351
    ab8448, ab69498, ab91655 and ab63856
    anti Von antibodies are;
    ab6994, ab11713 and ab9378 are the best antibodies available.
    Please click on the link to explore secondary antibodies with different conjugates;

    https://www.abcam.com/index.html?pageconfig=productmap&cl=918
    As per your email I think the application you are interested in, is double immunostaining. Please find attached the general protocols for double immunostaining.
    https://www.abcam.com/index.html?pageconfig=resource&rid=11458
    https://www.abcam.com/index.html?pageconfig=resource&rid=11459
    Double immunostaining can be divided in two types:
    Labelling using fluorochromes: The commonest fluorochrome available is FITC which gives green colour, TRITC and Texas red which gives red color. Other fluorochromes are Dylight 488 (Green), Dylight 633 (red) and so on. Please click the for complete list of fluorochromes (https://docs.abcam.com/pdf/immunology/fluorochromes_table_updated.pdf). The secondary antibodies conjugated to these fluorochromes can be used to detect 2 marker proteins same time in double immunolabelling. Please be advised that while selecting the fluorophores the absorption maxima of one dye should not overlap with other.
    Labelling using enzymatic methods: Enzyme labelled secondary antibodies are used in this method. The primary antibodies should either be from different species (e.g. rabbit or mouse) or both from same species but different in isotypes. When using secondary antibodies please use one HRP conjugated and other AP (alkaline phosphatase) conjugated ab so using substrate for each enzyme will give two colour staining.
    Things to remember in double staining
    - The primary antibodies should have different host
    - If the host is same they should be different isotypes e.g. IgG1a and IgG1 or IgG1 and IgG3 etc.
    - If the two targets are expressed on same part of cell then fluorochrome is best method, if the markers are expressed on different part of cells e.g. membrane or nucleus then enzymatic method is best.
    - Use the optimized dilution of primary antibodies.
    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
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Pig Tissue sections (Heart)
    Specification
    Heart
    Fixative
    Acetone
    Permeabilization
    Yes - 0.3 % Triton in PBS
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Dec 15 2011

    11-20 of 25 Abreviews or Q&A

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