The TGA kit is an assay system for determination of thrombin generation over time in platelet poor or platelet rich plasma (PPP or PRP) upon activation of the clotting cascade by micelles of negatively charged phospholipids containing different amounts of human tissue factor and CaCl2. The kit can be used to monitor hemophiliacs during inhibitor bypassing therapy, to monitor anticoagulation therapy, to calculate INR values for patients and to determine states of bleeding disorders or thrombophilia as well as the activity of circulating micro particles. This broad range of applications is possible by providing different tissue factor concentrations and by monitoring the whole kinetic of thrombin generation during initiation, amplification and down regulation of thrombin formation. TGA is therefore a universal assay kit for analyzing and monitoring the function of the haemostatic system.
Description :
TGA is based on monitoring the fluorescence generated by the cleavage of a fluorogenic substrate by thrombin over time upon activation of the coagulation cascade by different concentrations of tissue factor and negatively charged phospholipids in plasma. From the changes in fluorescence over time, the concentration of thrombin (nM) in the sample can be calculated using the respective thrombin calibration curve. The increase in thrombin concentration with time then allows to calculate generation of thrombin in the sample and to plot such thrombin values over time for the whole coagulation process. This then results in the visualization of the different phases of clot formation.
Storage :
Reagent RT*(20...25°C) +2…8°C -20°C TGA substrate (SUB) 1 week 1 month 6 months TGA buffer (BUF) 8 hours 1 week 1 month TGA thrombin calibrator (CAL) 8 hours 1 week 6 months TGA reagent B (RB), C Low (RCL) , High (RCH), D (RD) 8 hours 1 week 6 months TGA control high (CH) 4 hours 8 hours 1 month TGA control low (CL) 4 hours 8 hours 1 month Avoid contamination by micro-organisms. Plasmas should be frozen only once; during storage, the vials should be tightly capped. Stability of the sample material: * room temperature Sample material RT* (20...25°C) +2…8°C -20°C PPP, PRP and PFP Plasma 2 hours 4 hours 1 month An immediate centrifugation after blood withdrawal is recommended. Further we recommend an immediate shock freezing of the centrifuged samples. Attention! The frozen samples should be stored in a constant environment - avoid exposing the samples to variations in temperature. Before transportation we recommend to centrifuge and prepare the samples.
Size :
3x16 tests
Kit Components :
Substrate: 3 x 1.5 TGA substrate (SUB) Fluorogenic substrate 1 mM Z-G-G-RAMC, 15 mM CaCl2 Buffer: 1 x 3 TGA buffer (BUF) Hepes-NaCl-buffer containing 0.5 % bovine serum albumin Calibrator: 1 x 0.5 TGA thrombin calibrator ~1.000 nM thrombin in buffer with BSA Reagent C Low: 1 x 0.5 TGA reagent C (RC) Low RC Low conc. of phospholipid micelles containing rhTF in Tris-Hepes-NaCl buffer Reagent C High: 1 x 0.5 TGA reagent C (RC) High RC High conc. of phospholipid micelles containing rhTF in Tris-Hepes-NaCl buffer Reagent D: 1 x 1.5 TGA reagent D (RD) RD conc. of phospholipid micelles in TrisHepes-NaCl buffer Control High: 1 x 1 TGA control high (CH) Human plasma with increased thrombin generation, lyophilized. Control Low: 1 x 1 TGA control low (CL) Human plasma with decreased thrombin generation, lyophilized. Each reagent is available separately.
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