Pathromtin sl reagent

Manufactured by Siemens

Sourced in Austria

Pathromtin SL is a laboratory reagent used in the determination of the activated partial thromboplastin time (aPTT) for monitoring anticoagulant therapy and the investigation of coagulation disorders. It is a ready-to-use solution that contains phospholipids and ellagic acid, which activate the intrinsic pathway of the coagulation cascade.

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Lab products found in correlation

Thrombotrak solo coagulometer, by Axis-Shield (3 mentions) Multifibren u reagent, by Siemens (2 mentions) Thromborel s reagent, by Siemens (1 mentions) Cacl2, by Avantor (1 mentions) Prism 7, by GraphPad (1 mentions) Calcium chloride, by Thermo Fisher Scientific (1 mentions) Teg 5000 analyzer, by Haemonetics (1 mentions) Bcs xp coagulation analyzer, by Siemens (1 mentions) Innovance vwf ac assay, by Siemens (1 mentions)

Spelling variants of this product

Pathromtin SL (29 citations) Pathromtin (3 citations)

6 protocols using pathromtin sl reagent

Automated Hematology and Coagulation Assays

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The number of platelets was determined on the Sysmex XE-2100^TM Automated Hematology System (Sysmex Austria GmbH, Vienna, Austria). The PT (Thromborel^® S reagent, Siemens Healthcare Diagnostics GmbH, Vienna, Austria), APTT (Pathromtin^® SL reagent, Siemens Healthcare Diagnostics GmbH, Vienna, Austria), and fibrinogen (Multifibren^® U reagent, Siemens Healthcare Diagnostics GmbH, Vienna, Austria) were measured on the Siemens/Dade Behring BCS XP Analyzer Automated Coagulation System (Siemens Healthcare Diagnostics GmbH, Vienna, Austria). Daily QC measurements were performed with commercially available control material within the normal and pathological ranges (Control Plasma N and P (Siemens Healthcare Diagnostics GmbH, Vienna, Austria): PT 73.0 - 109.0% and 32 - 48%; APTT 28.0 - 38.0 s and 70 - 105 s; fibrinogen 2.2 - 3.2 g/L and 0.6 - 1.4 g/L, respectively; e-CHECK^®(XE) (Sysmex Austria GmbH): platelet counts 195 – 240 G/L, 30 – 75 G/L and 480 - 580 G/L). Inter-assay precision was calculated with results of minimum 20 consecutive days. One tenfold measurement was performed at one day to determine the intra-assay precision. The intra- and inter-assay CV were as follows: for PT 1.71 and 3.26%, respectively; for PT-INR 2.43 and 3.24%, respectively; for APTT 2.31 and 2.83%, respectively; for fibrinogen 1.23 and 2.87%, respectively; and for platelet count 1.23 and 3.4%, respectively.

Enko D., Mangge H., Münch A., Niedrist T., Mahla E., Metzler H, & Prüller F. (2017). Pneumatic tube system transport does not alter platelet function in optical and whole blood aggregometry, prothrombin time, activated partial thromboplastin time, platelet count and fibrinogen in patients on anti-platelet drug therapy. Biochemia Medica, 27(1), 217-224.

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Anticoagulant Activity of Sulphated Carbohydrates

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Serially diluted, sulphated carbohydrate samples (25 μl) were incubated with normal human citrated plasma (50 μl; NHSBT) and Pathromtin SL reagent (50 μl; Siemens) for 2 min at 37 C prior to the addition of calcium chloride (25 μl; 50 mM). The time taken for clot formations to occur (an upper maximal of 2 mins was observed) were recorded using a Thrombotrak Solo coagulometer (Axis-Shield) as per the manufacturer’s instructions. Water and sodium porcine mucosal heparin (203 IU/mg; VWR) were assayed as controls. The EC₅₀ values of all semi-synthetic, sulphated carbohydrates were determined using a sigmoidal dose response curved fitted post normalisation (with a 100% upper maximal at 2 mins; 0% lower maximal represented by the time required for the water control to clot normal human citrated plasma) with GraphPad Prism 6 software and compared to those obtained for the heparin control.

Skidmore M.A., Mustaffa K.M., Cooper L.C., Guimond S.E., Yates E.A, & Craig A.G. (2017). A semi-synthetic glycosaminoglycan analogue inhibits and reverses Plasmodium falciparum cytoadherence. PLoS ONE, 12(10), e0186276.

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Anticoagulation Assay for Glycosaminoglycans

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Glycosaminoglycan samples (or control), pooled, normal human plasma (citrated; Technoclone, Vienna, Austria) and Pathromtin SL reagent (Siemens, Munich, Germany) were incubated for 120 s (37 °C) prior to the addition of 50 mM CaCl₂ (VWR, Lutterworth, UK; Vtot = 175 uL, 1:2:2:2 v/v). Clot formation times were determined using a Thrombotrak Solo coagulometer (Axis-Shield, Dundee, UK). An upper maximal of 120 s, representing 100% inhibition of clotting was adopted. Water (0% clot inhibition, representing a normal aPTT clotting time, of around 37–40 s) and porcine heparin (193 IU.mg⁻¹; Celsus, Cincinnati, OH, USA) were used as controls. EC₅₀ values were calculated by the fitting of a sigmoidal dose response curve (GraphPad Prism 7, San Diego, CA, USA).

Mycroft-West C.J., Devlin A.J., Cooper L.C., Guimond S.E., Procter P., Guerrini M., Miller G.J., Fernig D.G., Yates E.A., Lima M.A, & Skidmore M.A. (2021). Glycosaminoglycans from Litopenaeus vannamei Inhibit the Alzheimer’s Disease β Secretase, BACE1. Marine Drugs, 19(4), 203.

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aPTT Clotting Time Inhibition Assay

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Serially diluted GAG samples (25 μL) were incubated with pooled, normal human citrated plasma (50 μL; Technoclone, Surrey, UK) and Pathromtin SL reagent (50 μL; Siemens, Erlangen, Germany) for 2 mins at 37 °C prior to the addition of calcium chloride (25 μL, 50 mM; Alfa Aesar, Heysham, UK). The time taken for clot formations to occur (an upper maximal of 2 mins was imposed, represented as 100% inhibition of clotting) was recorded using a Thrombotrak Solo coagulometer (Axis-Shield). HPLC-grade H₂O (0% inhibition of clotting, representing a normal aPTT clotting time, of ≈ 37–40 seconds) and porcine mucosal heparin (193 IU mg⁻¹; Celsus, Cincinnati, OH, USA) were screened as controls. The EC₅₀ values of all test and control samples were determined using a sigmoidal dose response curve fitted with Prism 7 (GraphPad Software, San Diego, CA, USA).

Mycroft-West C.J., Cooper L.C., Devlin A.J., Procter P., Guimond S.E., Guerrini M., Fernig D.G., Lima M.A., Yates E.A, & Skidmore M.A. (2019). A Glycosaminoglycan Extract from Portunus pelagicus Inhibits BACE1, the β Secretase Implicated in Alzheimer’s Disease. Marine Drugs, 17(5), 293.

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Comprehensive Coagulation Profile Analysis

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Fibrinogen (Multifibren^® U reagent, Siemens Healthcare Diagnostics GmbH, Vienna, Austria), Factor VIII (Pathromtin^® SL reagent and factor VIII deficient plasma, Siemens Healthcare Diagnostics GmbH, Vienna, Austria), Factor XIII (Berichrom FXIII reagent Siemens Healthcare Diagnostics GmbH, Vienna, Austria) as well as vWF Activity and ADAMTS13 (Innovance vWF Ac assay, Siemens Healthcare Diagnostics GmbH, Vienna, Austria) were measured on the Atellica COAG 360 Coagulation System (Siemens Healthcare Diagnostics GmbH, Vienna, Austria). Thrombin Generation was measured on the BCS XP Coagulation Analyzer using Endogenous Thrombin Reagent (Innovance ETP, (Siemens Healthcare Diagnostics GmbH, Vienna, Austria). Functional fibrinogen test was performed on the TEG 5000 analyzer (Haemonetics, Vienna, Austria) at the various time points, respectively.

Kovacic Krizanic K., Prüller F., Rosskopf K., Payrat J.M., Andresen S, & Schlenke P. (2022). Preparation and Storage of Cryoprecipitate Derived from Amotosalen and UVA-Treated Apheresis Plasma and Assessment of In Vitro Quality Parameters. Pathogens, 11(7), 805.

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aPTT Assay for Intrinsic Coagulation

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Activated partial thromboplastin time (aPTT) was measured using a Kugelkoagulometer (ABW Medizin und Technik GmbH)^{7 (link)}. Briefly, 50 µL of FXII-deficient human or murine plasma, reconstituted to physiologic FXII levels (375 nM) with FXII_pd or recombinant FXII proteins or buffer control (20 µL), was added to 50 µL Pathromtin SL reagent (Siemens HealthCare Diagnostics). After incubation for 120 s at 37 °C, 30 μL of CaCl₂ (62.5 mM) was added and clotting times were recorded. For the clotting assay with pAb-αGAL14, normal PPP was pre-incubated with buffer, insoluble polyP (10 μg/mL), pAb-αGAL14 (end concentration of 2 μM) or an antibody against VASP (pAb-αControl)^{68 (link)} before recalcification. For the clotting assay with mAb-αPR-III_37, 50 µL normal PPP was spiked with buffer or mAb-αPR-III_37, in the presence of various concentrations of long-chain polyP followed by recalcification. The Kugelkoagulometer aPTT assays have limited sensitivity at and above a system-specific threshold of 180 s.

Heestermans M., Naudin C., Mailer R.K., Konrath S., Klaetschke K., Jämsä A., Frye M., Deppermann C., Pula G., Kuta P., Friese M.A., Gelderblom M., Sickmann A., Preston R.J., Nofer J.R., Rose-John S., Butler L.M., Salomon O., Stavrou E.X, & Renné T. (2021). Identification of the factor XII contact activation site enables sensitive coagulation diagnostics. Nature Communications, 12, 5596.

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