Horseradish peroxidase conjugated anti human vwf antibodies

Manufactured by Agilent Technologies

Sourced in Denmark

Horseradish peroxidase conjugated anti-human VWF antibodies are laboratory reagents used for the detection and quantification of human von Willebrand factor (VWF) in various analytical techniques. These antibodies are conjugated with the enzyme horseradish peroxidase, which enables signal amplification and sensitive measurement of target analytes.

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Lab products found in correlation

Microplate reader, by Agilent Technologies (1 mentions) Acl 300, by Werfen (1 mentions) Anti mpo monoclonal antibody clone 4a4, by Bio-Rad (1 mentions) Synergy ht, by Agilent Technologies (1 mentions) Cell death detection elisa kit, by Roche (1 mentions) Cell death detection elisaplus, by Roche (1 mentions)

3 protocols using horseradish peroxidase conjugated anti human vwf antibodies

Quantifying VWF and ADAMTS13 Levels

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VWF antigen (VWF:Ag) was determined with an in-house ELISA with polyclonal rabbit anti-human VWF antibodies and horseradish peroxidase conjugated anti-human VWF antibodies (DakoCytomation, Glostrop, Denmark) for catching and tagging, respectively.
VWF collagen binding (VWF:CB) was determined with an in-house ELISA using collagen bovine antibodies and horseradish peroxidase conjugated anti-human VWF antibodies (DakoCytomation, Glostrop, Denmark) for catching and tagging, respectively.
ADAMTS13 activity was measured by a Fluorescence Resonance Energy Transfer Substrate (FRETS) assay using a fluorescent VWF peptide consisting of 73 amino acids (FRETS-VWF73). Plates were read with BioTek’s microplate reader (BioTek). The intra-assay coefficients of variation for VWF:Ag, VWF:CB and ADAMTS13 were 1.9%, 4.5% and 3.6%, respectively. The inter-assay coefficients of variation for VWF:Ag, VWF:CB and ADAMTS13 were 8.3%, 7.9% and 7.5%, respectively.

van Loon J.E., Sonneveld M.A., Praet S.F., de Maat M.P, & Leebeek F.W. (2014). Performance Related Factors Are the Main Determinants of the von Willebrand Factor Response to Exhaustive Physical Exercise. PLoS ONE, 9(3), e91687.

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Quantifying Hemostasis Biomarkers in Plasma

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Fibrinogen levels were derived from the clotting curve of the prothrombin time assay, using Thromborel S (Behringwerke, Marburg, Germany) on the ACL 300 coagulation analyzer (Instrumentation Laboratory). vWF:Ag levels were measured with an in-house ELISA using polyclonal rabbit antihuman VWF antibodies and horseradish-peroxidase-conjugated antihuman VWF antibodies (DakoCytomation, Glostrup, Denmark) to catch and tag vWF. ADAMTS13 activity was measured in a kinetic assay using Fluorescence Resonance Energy Transfer Substrate VWF 73 (FRETS-VWF73), as is thoroughly described in the previous articles [16 (link), 17 (link)].
We determined NET levels by measuring MPO–DNA complexes with an ELISA as reported earlier [18 (link)]. We adjusted the commercial human cell death ELISA kit (Cell death detection ELISAPLUS, Roche Diagnostics Nederland B.V., Almere, The Netherlands). Briefly, as the capturing antibody, we used anti-MPO monoclonal antibody (clone 4A4, ABD Serotec). Patient plasma was added in combination with the peroxidase-labeled anti-DNA monoclonal antibody (from cell death detection ELISA kit; Roche). The absorbance at 405 nm wavelength was measured using Biotek Synergy HT plate reader with a reference filter of 490 nm. The values are expressed as milli-arbitrary units (mAU/mL).

Tilly M.J., Geurts S., Donkel S.J., Ikram M.A., de Groot N.M., de Maat M.P, & Kavousi M. (2021). Immunothrombosis and new-onset atrial fibrillation in the general population: the Rotterdam Study. Clinical Research in Cardiology, 111(1), 96-104.

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Measurement of von Willebrand Factor

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At inclusion in the study, venous blood was collected in 0.105 M sodium citrate tubes (1 : 10) and centrifuged twice at 2200 9 g for 10 min at room temperature; plasma was stored at À 80 °C. Plasma levels of VWF:Ag and VWF activity (VWF:Act) were measured centrally (Erasmus University Medical Center, Rotterdam). VWF: Ag levels were determined with an in-house ELISA using polyclonal rabbit anti-human VWF antibodies and horseradish peroxidase-conjugated anti-human VWF antibodies (DakoCytomation, Glostrup, Denmark) for detection. VWF:Act was assessed with a LIA test, which uses mAbs directed against the glycoprotein (GP)Iba-binding domain of VWF and thereby reflects the binding of VWF to GPIba (HemosIL von Willebrand Factor Activity; Instrumentation Laboratory, Breda, the Netherlands). Phenotypic blood group was determined by mixing plasma from patients with red blood cells from donors with a known blood group [25] . If the phenotypic blood group was unknown, blood group was determined by genotyping the ABO blood group-specific SNPs rs687289 (marker for blood group O), rs507666 (marker for blood group A1), rs8176704 (marker for blood group A2), and rs8176749 (marker for blood group B) [26] . Details of the blood-sampling procedure and laboratory measurements at inclusion in the study have been described in more detail elsewhere [2] .

Sanders Y.V., van der Bom J.G., Isaacs A., Cnossen M.H., de Maat M.P., Laros-van Gorkom B.A., Fijnvandraat K., Meijer K., van Duijn C.M., Mauser-Bunschoten E.P., Eikenboom J, & Leebeek F.W. (2015). CLEC4M and STXBP5 gene variations contribute to von Willebrand factor level variation in von Willebrand disease. Journal of thrombosis and haemostasis : JTH, 13(6).

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