Plasma from 4 healthy donors was pooled and used for aPTT measurements. All measurements were performed at 37.2 °C using the MC-10 coagulometer (Merlin Medical, Tredegar, United Kingdom). In short, 50 µL of plasma was warmed up in the cuvette for 2 minutes, then 50 µL of undiluted Dade Actin FS (Siemens Healthcare Diagnostics, Marburg, Germany) was added and after another 2 minutes 50 µL of 25 mM CaCl2 was added and clotting time was measured.
Dade actin fs
Manufactured by Siemens
Sourced in Germany
The Dade Actin FS is a laboratory equipment product manufactured by Siemens. It is designed for the quantitative determination of fibrin/fibrinogen degradation products (FDP) in human plasma. The Dade Actin FS utilizes an immunoagglutination method to measure FDP levels.
Automatically generated - may contain errors
Lab products found in correlation
Kaolin, by Merck Group (1 mentions)
Rha infestin 4, by CSL Behring (1 mentions)
Sta c k prest, by Diagnostica Stago (1 mentions)
Dornase alfa, by Roche (1 mentions)
Mp reagent, by Diagnostica Stago (1 mentions)
Pathromtin sl, by Siemens (1 mentions)
Heparin, by Merck Group (1 mentions)
Cs2100i, by Sysmex (1 mentions)
Cs 2500, by Sysmex (1 mentions)
Thromboclotin, by Siemens (1 mentions)
Dade innovin, by Siemens (1 mentions)
Dade thrombin reagent, by Siemens (1 mentions)
Atellica coag 360, by Siemens (1 mentions)
Vacutainer tube, by BD (1 mentions)
Sta r evolution analyzer, by Diagnostica Stago (1 mentions)
Hemoclot, by Hyphen Biomed (1 mentions)
Sta compact analyzer, by Diagnostica Stago (1 mentions)
Thromborel s, by Siemens (1 mentions)
8 protocols using dade actin fs
1
Activated Partial Thromboplastin Time Assay
2
Evaluating Polyphosphate Effects on Coagulation
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Kaolin was obtained from Sigma Aldrich. The activated partial thromboplastin time (aPTT) reagent Dade® Actin® FS containing ellagic acid and soybean phospholipids and Pathromtin SL® were purchased from Siemens Healthcare Diagnostics. FXII contact activators STA® - C.K. Prest® and STA® - Cephascreen® were obtained from Diagnostica Stago (Asnières, France), while SynthAFax®, SynthASil® and APTT SP reagent were purchased from IL Werfen (München, Germany). The following synthetic polyphosphates with different lengths were obtained from ICL Pharmaceuticals: ammonium polyphosphate (P42, #2 2846), ammonium polyphosphate (P30, #2 2840), sodium polyphosphate (P75, #7 9990), sodium polyphosphate (P70, #7 1480), sodium polyphosphate (P68, #7 1480). Dornase alfa, the recombinant human DNaseI (Pulmozyme®), was purchased from Roche. FXII purified from human plasma was obtained from Haematologic Technologies. The FXIIa-specific inhibitor rHA-Infestin-4 was supplied by CSL Behring. Thrombin generation assays were performed with the fluorogenic substrate Z-Gly-Gly-Arg-AMC, purchased from Bachem Bubendorf, Switzerland. The thrombin calibrator (α2-macroglobulin-thrombin complex, α2M-T) and MP reagent (containing phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine at a ratio of 20:60:20) were obtained from Diagnostica Stago.
3
Anticoagulant Polymer Effects on Clotting Times
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The blood obtained from healthy donors was collected into sodium citrate 3.2% tubes. Platelet poor plasma (PPP) was obtained by centrifuging the blood at 3100g for 25 min at 20 °C. The polymers were dissolved in PBS buffer containing 5% DMSO. Polymers in the solution were mixed with PPP in volume:volume ratio 1:9 resulting in the final polymers concentration of 10 and 100 mg/L. Prothrombin time was measured using MRX Owren's PT reagent (MediRox, GHI 131-10) and activated partial thromboplastin time was measured using Dade Actin FS (Siemens, B4218-100). All measurements were performed on Sysmex CS-2100i. PBS and PBS with 5% DMSO were used as a negative control. Heparin (Sigma, H3393) was used as a positive control.
4
FVIII and VWF Measurement Protocols
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FVIII:C was measured using the one-stage assay with ACL 9000TM (Werfenmedical IL, Bedford, MA, USA) using Synthasil® reagent or CS-2500 automated coagulation analyser (Sysmex Corporation, Kobe, Japan) using the Dade Actin FS (Siemens Healthcare Diagnostics, Marburg, Germany) reagent. VWF:Ag was measured with an ACL 9000TM automated coagulation analyser using HemosILTM VWF antigen (Werfenmedical IL, Bedford, MA, USA).
5
Anticoagulant Effects of Fasciola Serpins
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Calibrated automated thrombography (CAT) was used to assess the interaction of the F. hepatica serpins with coagulation factors II, V, VII, VIII, IX, X and XI as per the protocol used by Huskens et al. [40 (link)]. Thrombin generation was tested in triplicate in the presence or absence of rFhSrp1 and rFhSrp2, which were added in varying concentrations (12.5 to 50 μg/mL).
The inhibitory potential of the F. hepatica serpins against the intrinsic coagulation pathway was investigated using the activated partial thromboplastin time (APTT) test. The tests were performed in triplicate using varying concentrations of the F. hepatica serpins (12.5 to 50 μg/mL), at 37.2°C using the MC-10 coagulometer (Merlin Medical, Tredegar, UK). Plasma, with or without the recombinant F. hepatica serpins, was pre-incubated at 37.2°C, followed by the addition of 50 μL of undiluted Dade Actin FS (Siemens Healthcare Diagnostics). After 2 min incubation, 50 μL of 25 mM CaCl2 was added and the clotting time measured.
The inhibitory potential of the F. hepatica serpins against the intrinsic coagulation pathway was investigated using the activated partial thromboplastin time (APTT) test. The tests were performed in triplicate using varying concentrations of the F. hepatica serpins (12.5 to 50 μg/mL), at 37.2°C using the MC-10 coagulometer (Merlin Medical, Tredegar, UK). Plasma, with or without the recombinant F. hepatica serpins, was pre-incubated at 37.2°C, followed by the addition of 50 μL of undiluted Dade Actin FS (Siemens Healthcare Diagnostics). After 2 min incubation, 50 μL of 25 mM CaCl2 was added and the clotting time measured.
6
Coagulation Factors Quantification Protocol
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The Clauss fibrinogen tests (Dade Thrombin Reagent, Siemens, Marburg, Germany), PT (Dade Innovin, Siemens, Marburg, Germany), APTT (Dade Actin FS, Siemens, Marburg, Germany) and TT (Thromboclotin, Siemens, Marburg, Germany) were processed using a Sysmex CS-5100 haematology analyser. The normal range for each test is as follows: fibrinogen (1.5–4.5 g/L), PT (13–16 s), APTT (26–36 s) and TT (16–19 s). Coagulation factors II, V, VII, VIII, IX, X, XI and XII were quantified using a Siemens Sysmex CS-5100 haematology analyser. FXIII and vWF were measured using a Siemens Sysmex CS-5100 haematology analyser and Berichrom Factor XIII Chromogenic (Siemens, Marburg, Germany) and vWF:Ag assay kit (Siemens, Marburg, Germany), respectively. Normal ranges for all coagulation factors are 50–150%. α2AP concentration was measured using an in-house ELISA, respectively.
7
Coagulation Marker Evaluation Protocol
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All parameters were measured in the central laboratory of the university Hospital of Tuebingen on an Atellica COAG 360 coagulation platform (Siemens Healthcare Diagnostics, Marburg, Germany) as reported elsewhere. 22 The following reagents from Siemens Healthcare Diagnostics, Marburg, Germany were used: Dade Actin FS for activated partial thromboplastin time (aPTT), INNOVANCE D-dimer assay for D-dimer, and Dade thrombin for fibrinogen.
8
Plasma Coagulation Tests for Dabigatran
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Blood was collected into a Vacutainer tube (Becton Dickinson Canada, Mississauga, Ontario, Canada) containing sodium citrate (0.105 M, 3.2%), centrifuged for 15 min at 1500 9g, and double-spun to remove platelets. Platelet-poor plasma was separated into aliquots and stored at À 70 °C. Plasma samples were analyzed at the Hamilton Regional Special Coagulation Laboratory by personnel who were blinded to patient characteristics.
The coagulation function tests (and assays) performed were: PT (Thromborel S; Siemens, Marburg, Germany); APTT (Dade Actin FS; Siemens ); TT (Sigma-Aldrich, Oakville, Ontario, Canada); and dTT (Hemoclot; Hyphen BioMed, Neuville-sur-Oise, France). The APTT and TT were measured on am STA Compact analyzer, and the dTT was measured on a STAR Evolution analyzer (both from Diagnostica Stago, Asni eres-sur-Seine, France). Plasma dabigatran levels were measured with a validated HPLC method (AAI Pharma Deutschland, Neu-Ulm, Germany) [30] . The results from these coagulation tests were not available for clinical use, but study participants could have clinically indicated laboratory testing, including coagulation tests.
The coagulation function tests (and assays) performed were: PT (Thromborel S; Siemens, Marburg, Germany); APTT (Dade Actin FS; Siemens ); TT (Sigma-Aldrich, Oakville, Ontario, Canada); and dTT (Hemoclot; Hyphen BioMed, Neuville-sur-Oise, France). The APTT and TT were measured on am STA Compact analyzer, and the dTT was measured on a STAR Evolution analyzer (both from Diagnostica Stago, Asni eres-sur-Seine, France). Plasma dabigatran levels were measured with a validated HPLC method (AAI Pharma Deutschland, Neu-Ulm, Germany) [30] . The results from these coagulation tests were not available for clinical use, but study participants could have clinically indicated laboratory testing, including coagulation tests.
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