Overview

  • Product name
    Mouse Von Willebrand Factor A2 ELISA Kit, Fluorescent
    See all Von Willebrand Factor kits
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Plasma 5 5.3%
    Inter-assay
    Sample n Mean SD CV%
    Plasma 3 13.3%
  • Sample type
    Cell culture supernatant, Serum, Heparin Plasma, EDTA Plasma, Citrate Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    7.1 pg/ml
  • Range
    13.67 pg/ml - 14000 pg/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 83.6 80.2% - 89.7%
    Heparin Plasma 82.2 78.8% - 86.7%
    EDTA Plasma 94.4 88.2% - 98.3%
    Citrate Plasma 86.3 82.7% - 90.3%
    serum free media 79 70.9% - 83.8%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Mouse
    Does not react with: Rabbit, Goat, Dog, Human, Pig
  • Product overview

    Von Willebrand Factor A2 in vitro CatchPoint® SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Von Willebrand Factor A2 protein in mouse serum, plasma, and cell culture supernatant samples.


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint® 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.  CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    vWF A2 and vWF (von Willebrand Factor) are distinct proteins with non-overlapping amino acid sequences generated by proteolytic cleavage from the same precursor protein during intracellular processing. vWF is important in the maintenance of hemostasis; it is a glycoprotein that circulates in plasma as a series of high molecular weight multimers and mediates the adhesion of platelets to exposed sub-endothelium. vWF is synthesized in endothelial cells and megakaryocytes. vWF A2 is found in plasma and platelets, from which they are released by their activation including thrombin. Both vWF and vWF A2 are deficient in von Willebrand’s disease.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    100X Stoplight Red Substrate 1 x 120µl
    10X Mouse vWF A2 Capture Antibody 1 x 600µl
    10X Mouse vWF A2 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent 4BR 1 x 6ml
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
    Mouse vWF A2 Lyophilized Recombinant Protein 2 vials
  • 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
    • VWD
    • vWF
    • VWF_HUMAN
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229397 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

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

  • The concentrations of vWF A2 were measured in duplicates, interpolated from the vWF A2 standard curves and corrected for sample dilution. Undiluted samples are as follows: serum 0.75 %, plasma (citrate) 3.5 %, plasma (EDTA) 7.0 %, plasma (heparin) 3.5 %. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean vWF A2 concentration was determined to be 1003 ng/mL in serum, 146 ng/mL in plasma (citrate) 118 ng/mL in plasma (EDTA) and 190 ng/mL in plasma (heparin).

  • The concentrations of vWF A2 were measured in duplicates, interpolated from the vWF A2 standard curves and corrected for sample dilution. Undiluted samples are as follows: 7,000 pg/mL of vWF A2 in 100% serum-free cell culture media. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean spiked vWF A2 concentration was determined to be 5962 pg/mL in serum-free cell culture media.

Protocols

References

ab229397 has not yet been referenced specifically in any publications.

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