Mouse Von Willebrand Factor A2 ELISA Kit, Fluorescent (ab229397)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 7.1 pg/ml
- Range: 13.67 pg/ml - 14000 pg/ml
- Sample type: Cell culture supernatant, Cit plasma, EDTA Plasma, Hep Plasma, Serum
- Detection method: Fluorescent
- Assay type: Sandwich (quantitative)
- Reacts with: Mouse
Overview
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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, Hep Plasma, EDTA Plasma, Cit 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% Hep Plasma 82.2 78.8% - 86.7% EDTA Plasma 94.4 88.2% - 98.3% Cit 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: Goat, 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.
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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.
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Platform
Pre-coated microplate (12 x 8 well strips)
Properties
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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 (ab193972) 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
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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
modificationsAll 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
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Alternative names
- Coagulation factor VIII
- Coagulation factor VIII VWF
- F8VWF
see all -
Database links
- Entrez Gene: 22371 Mouse
- SwissProt: Q8CIZ8 Mouse
- Unigene: 22339 Mouse
Images
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Other - Mouse Von Willebrand Factor A2 ELISA Kit, Fluorescent (ab229397)
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.
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Example of mouse Von Willebrand Factor A2 standard curve in Sample Diluent NSBackground-subtracted data values (mean +/- SD) are graphed.
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Interpolated concentrations of native vWF A2 in mouse serum and plasma samples.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).
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Interpolated concentrations of spiked mouse recombinant vWF A2 in serum-free cell culture media samples.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.
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Sandwich ELISA - Mouse Von Willebrand Factor A2 ELISA Kit, Fluorescent (ab229397)To learn more about the advantages of recombinant antibodies see here.
