Z007 zedichrom, fxiiiassaykit

Assay for the determination of Factor XIII activity Blood (9 vol.) must be col ected on 0.109 M citrate anticoagulant (1 vol.); Determination of Factor XIII activity in plasma samples. plasma supernatant is decanted fol owing a 10 min centrifugation at 2,500 g; citrated plasma should be tested within 8 hours or stored frozen at -20 °C or Factor XIII is converted by Thrombin into Factor XIIIa. At the same time, Thrombin converts Fibrinogen into Fibrin. The clotting is prevented by an Prewarm the REAGENT MIXTURE to 37 °C before testing. Add 100 µl of Factor XIIIa is a transglutaminase incorporating glycine methyl ester (amine sample to 1,000 µl of REAGENT MIXTURE, mix thoroughly and start donor) into a peptide substrate (glutamine donor). measuring at 37 °C at the fol owing instrument parameters: The concomitant release of ammonia is monitored by an enzymatic reaction consuming NADPH. The decrease of NADPH is determined at 340nm. (1) 3 X ACTIVATOR REAGENT (add 5mL water each) (2) 3 x NADPH REAGENT (dissolve in 5mL ACTIVATOR REAGENT each) (3) 3 X DETECTION REAGENT (add 5mL water each) human thrombin, clot inhibitor peptide, calcium chloride, heparin antagonist (hexadimethrine bromide), albumin (bovine), buffer salts and sodium azide The NADPH REAGENT contains NADPH, albumin (bovine) and sodium azide (preservative). glutamine donor substrate peptide (FXIIIa substrate), glycine methyl ester The decrease in absorption after reaching the linear phase (typical y period (amine donor), GLDH (glutamate dehydrogenase), ADP (adenosine between 7 and 12 minutes after starting the reaction by adding the sample) is proportional to the Factor XIII activity. α-ketoglutarate (GLDH substrate), albumin (bovine), buffer salts and sodium azide (preservative). The results can be evaluated using a reference curve. To calculate the reference curve, Standard Human Plasma is diluted with saline and Reagent preparation, storage and stability In their original packing box, when stored at 2-8 °C, the unopened reagents are stable until the expiration date printed on the box. Dissolve one vial of ACTIVATOR REAGENT (1) in 5 mL of distil ed water. The use of the ZEDICHROM FACTOR XIII ASSAY should be careful y validated Transfer the whole volume to one vial of NADPH REAGENT (2). by using suitable CONTROL PLASMAS. Basical y, the assay can be performed in a 96 wel plate reader. However, it should be noted that the assay is Dissolve one vial of DETECTION REAGENT (3) in 5 mL of distil ed water. meant for research and development only. It can be feasible varying the sample to REAGENT MIXTURE to improve results. Combine both solutions (10mL) and mix thoroughly (REAGENT MIXTURE). Very low or very high concentrations of fibrinogen could influence the Stability of the REAGENT MIXTURE after reconstitution Reference Range, Precision and Specificity The range has been found to be 70 – 140 % of normal. The coefficient of variation in the series was 3.6% for STANDARD HUMAN PLASMA. Day to day variance was 3.6%. No interfering plasma activities are known to the 1) Fickenscher K, Aab A, Stüber W.A; photometric assay for blood coagulation factor XIII. Thromb Haemost. 1991 May 6;65(5):535-40. 2) Levente Kárpáti et al.; A Modified, Optimized Kinetic Photometric Assay for the The ZediChrom assay can be used in standard UV spectrophotometers. Determination of Blood Coagulation Factor XIII Activity in Plasma. Clinical Chemistry. Refer to the instructions of the manufacturer. Verwendete Symbole / Used symbols / Använda symboler / Anvendte symboler

Source: http://www.zedira.eu/resources/content/pdf/z007.pdf

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