Kc4 coagulation analyzer

Manufactured by Trinity Biotech

Sourced in Ireland

The KC4 coagulation analyzer is a laboratory instrument used for the measurement and analysis of coagulation parameters in blood samples. It provides accurate and reliable data to support clinical decision-making in the diagnosis and monitoring of coagulation disorders.

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Lab products found in correlation

Accucheck fluorescent beads, by Thermo Fisher Scientific (1 mentions) Dade innovin, by Siemens (1 mentions) Facs lysing solution, by BD (1 mentions) Lsr 2 flow cytometer, by BD (1 mentions)

5 protocols using kc4 coagulation analyzer

Monoclonal Antibody Inhibition of FXI

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Derivation and activity of the murine anti-mouse FXI monoclonal antibody, 14E11, used in this study have been described elsewhere.^{35 (link)} Briefly, the antibody was generated by immunizing FXI-deficient mice with recombinant mouse FXI. Clone 14E11 was expanded in a CL1000 bioreactor (Integra Biosciences) and immunoglobulin G (IgG) was purified by cation exchange and thiophilic agarose chromatography.
Dosage of 14E11 for this study to achieve its maximal effect on prolonged activated partial thromboplastin time (APTT), used as a marker of pharmacological inhibition of the contact system activation over the course of the study, was determined based on a dose-finding experiment wherein C57BL/6 mice were injected with a single subcutaneous (s.c.) dose of 14E11 (4 mg/kg) and APTT was monitored over 10 days. Whole blood was collected into sodium citrate (0.32% w/v) at days 0, 3, 6, and 10 postinjection. Platelet-poor plasma (PPP) was isolated by centrifuging whole blood at 2000g for 10 min at room temperature (RT). PPP was mixed 1:1 with APTT reagent and incubated for 3 min at 37°C. CaCl₂ (8.3 mM final) was added in equal volume to PPP and APTT reagent and time to clot formation was measured using KC4 coagulation analyzer (Trinity Biotech). Throughout the study, plasma was serially collected from mice to analyze the FXI levels following the administration of 14E11.

Ngo A.T., Jordan K.R., Mueller P.A., Hagen M.W., Reitsma S.E., Puy C., Revenko A.S., Lorentz C.U., Tucker E.I., Cheng Q., Hinds M.T., Fazio S., Monia B.P., Gailani D., Gruber A., Tavori H, & McCarty O.J. (2021). Pharmacological targeting of coagulation factor XI mitigates the development of experimental atherosclerosis in low-density lipoprotein receptor-deficient mice. Journal of thrombosis and haemostasis : JTH, 19(4), 1001-1017.

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Blood Collection and Plasma Coagulation Assay

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Whole blood from euthanized mice was collected by venipuncture directly into sodium citrate (3.2% w/v) at a ratio of 9:1 v/v. Platelet poor plasma was obtained from citrated blood after centrifugation and stored in −80^o C.^{17 (link)} Thawed plasma was recalcified and protimes measured on a KC4 coagulation analyzer (Trinity Biotech) after recalcification to 8.3 mM and mixing with thromboplastin (Dade Innovin®, Siemens Healthcare Diagnostics).
Quantification of antigen-specific cell numbers in peripheral blood was measured using a whole blood assay.^{18 (link)} Blood was stained directly with fluorescent antibody cocktails along with PE-conjugated Kb- SIYRYYGL pentamers. AccuCheck fluorescent beads (Invitrogen) were added to each sample before lysing the red blood cells with BD FACS lysing solution (BD Biosciences) and samples analyzed on a BD LSRII flow cytometer. Cell concentrations were determined by comparing cellular events to bead events.

Tormoen G.W., Blair T.C., Bambina S., Kramer G., Baird J., Rahmani R., Holland J.M., McCarty O.J., Baine M.J., Verma V., Nabavizadeh N., Gough M.J, & Crittenden M. (2020). Targeting MerTK enhances adaptive immune responses following radiotherapy. International journal of radiation oncology, biology, physics, 108(1), 93-103.

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Plasma Clotting Time Measurement

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Human platelet-poor plasma (PPP) was prepared and clotting time measured as previously described [4 (link)]. Plasma (50 μl) was incubated with HBS, or 5 μM SCP (50 μl) in HBS, for 3 min at 37°C in coagulometer cuvettes. Subsequently, 50 μl of CaCl₂ (8.3 mM final) and diluted Innovin (8 pM final TF concentration) were added, and time to clot formation was measured with a KC4 Coagulation Analyzer (Trinity Biotech, Ireland). In selected experiments, FXI- or FIX-depleted PPP was used.

Puy C., Tucker E.I., Ivanov I.S., Gailani D., Smith S.A., Morrissey J.H., Gruber A, & McCarty O.J. (2016). Platelet-Derived Short-Chain Polyphosphates Enhance the Inactivation of Tissue Factor Pathway Inhibitor by Activated Coagulation Factor XI. PLoS ONE, 11(10), e0165172.

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Coagulation Changes by Biomaterial Carriers

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To evaluate changes in the coagulation properties of blood as a result of contact with carriers, we measured such coagulation indicators as prothrombin time (PT, an indicator of the external coagulation pathway) and activated partial thromboplastin time (APTT, an indicator of the internal coagulation pathway). These were measured with an Amelung KC4 coagulation analyzer (TRINITY Biotech, Ireland) in duplicate. Blood from four healthy donors was dispensed into tubes with sodium citrate as the anticoagulant and was incubated with particles (MNPs(BSA–TA) and BSA–TA). The scheme of the experiment was the same as described by Dobrovolskaia et al. [29 (link)]. A solution of 0.9% NaCl was used as the reference. After incubation, plasma was isolated by collecting the supernatant liquid and was subjected to clotting tests.

Novoselova M.V., German S.V., Sindeeva O.A., Kulikov O.A., Minaeva O.V., Brodovskaya E.P., Ageev V.P., Zharkov M.N., Pyataev N.A., Sukhorukov G.B, & Gorin D.A. (2019). Submicron-Sized Nanocomposite Magnetic-Sensitive Carriers: Controllable Organ Distribution and Biological Effects. Polymers, 11(6), 1082.

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Activated Partial Thromboplastin Time Assay

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Whole blood was collected from a single direct puncture of inferior vena cava after midline laparotomy into sodium citrate (0.32% w/v). Platelet-poor plasma (PPP) was isolated by centrifuging blood samples at 2000 g for 10 min at room temperature. PPP was mixed 1:1 with aPTT reagent and incubated for 3 min at 37°C. CaCl2 (8.3 mM final concentration) was added in equal volume to PPP and aPTT reagent. The time (seconds) to thrombus formation was measured using a KC4 coagulation analyzer (Trinity Biotech, Jamestown, NY).

Jordan K.R., Wyatt C.R., Fallon M.E., Woltjer R., Neuwelt E.A., Cheng Q., Gailani D., Lorentz C., Tucker E.I., McCarty O.J., Hinds M.T, & Nguyen K.P. (2022). Pharmacological reduction of coagulation factor XI reduces macrophage accumulation and accelerates deep vein thrombosis resolution in a mouse model of venous thrombosis. Journal of thrombosis and haemostasis : JTH, 20(9), 2035-2045.

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