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Plasmin activity in the pleural washes of WT and PAI-1−/− mice treated with saline or S. pneumoniae was measured by amidolytic assay using a plasmin substrate PL-52688, CeNorwalkm InCT Nonwalk CT) on a SpectraMax 96-well optical absorbance plate reader (Molecular devices, Sunnyvale, CA), as previously described [14].
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It was found to be capable of cleaving a plasmin substrate, further supporting the participation of rhinocerase in fibrinolysis (table 2).
As a consequence, the new display is 90% lighter than comparable ones using a glass substrate.
Also, consider using a different substrate.
With the ability to detect alterations in heat using a substrate underlay, the patch changes color when it senses an issue.
Our assays showed that when using an amidolytic chromophogenic substrate, the K m value of rhμPlg was similar to that of plasmin (Fig. 4).
Do not use a dirt based substrate.
It binds plasmin using a C-terminal lysine-rich domain and inhibits it by forming a stable, covalent serpin-enzyme complex [ 20].
Fibrinolytic activity was also analysed with a fluorescent plasmin substrate [Ala-Leu-Lys-AMC (D-Ala-Leu-Lys-7-amido-4-methylcoumarin) A8171, from Sigma Aldrich, Poole, UK].
IIa, uPA, and the His6 tag were separated from μ-plasmin using a Superdex-200 gel-filtration column.
Plasmin-plasmin inhibitor complexes (PPIC, PAP) were measured using a 96-well microtiter plate ELISA from DRG Instruments GmbH, Marburg, Germany.
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