Overview

  • Product name

    Human Von Willebrand Factor ELISA Kit
    See all Von Willebrand Factor kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Serum 3 3.4%
    Inter-assay
    Sample n Mean SD CV%
    Serum 5 6.4%
  • Sample type

    Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.079 ng/ml
  • Range

    0.469 ng/ml - 30 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 101 94% - 107%
    Cell culture media 88 80% - 95%
    Heparin Plasma 95 89% - 102%
    EDTA Plasma 92 86% - 97%
    Citrate Plasma 98 83% - 118%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Human, Rhesus monkey
    Does not react with: Mouse, Rat, Cow
  • Product overview

    Human Von Willebrand Factor SimpleStep ELISA® kit (ab189571) has been re-developed with new capture and detector antibodies. This new kit has the same name but a different product number (ab223864). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantifying Von Willebrand Factor in human serum, plasma and cell culture supernatant samples.


    Von Willebrand Factor in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Von Willebrand Factor protein in  human serum, plasma and cell culture supernatant samples.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    Von Willebrand Factor is critical 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. Von Willebrand Factor 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.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT (ab206977) 1 x 20ml
    Antibody Diluent 5BI 1 x 6ml
    Human Von Willabrand Factor Capture Antibody (Lyophilized) 1 vial
    10X Human Von Willabrand Factor Detector Antibody 1 x 600µl
    Human Von Willabrand Factor Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas

  • 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
  • Alternative names

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

Applications

Our Abpromise guarantee covers the use of ab223864 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Von Willebrand Factor were measured in duplicates, interpolated from the Von Willebrand Factor standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 1:500, plasma (citrate) 1:500, plasma (heparin) 1:500, and plasma (EDTA) 1:500. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Von Willebrand Factor concentration was determined to be 4.226 µg/mL in neat serum, 2.793 µg/mL in neat plasma (citrate), 1.774 µg/mL in neat plasma (heparin), and 2.819 µg/mL in neat plasma (EDTA).

  • Interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Von Willebrand Factor concentration was determined to be 5.280 µg/mL with a range of 2.088 – 11.35 µg/mL.

Protocols

References

ab223864 has not yet been referenced specifically in any publications.

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