The turbidity and turbidity-lysis protocols which have previously been used in patient samples [7 (link)] were adapted for use in murine plasma. Murine plasma samples (100μl, diluted 1:4 in 0.05mol/L HEPES buffer with 0.1% BSA, pH 7.4) were added to each well of a 96-well Greiner bio-one microtitre plate. An activation mix (25μl) containing murine thrombin (MCT-5020-50, Cambridge Biosciences, UK), calcium chloride (CaCl2), with or without human recombinant tissue plasmingen activator (tPA) in a 0.05mol/L HEPES buffer, were added to each well. Final concentrations per well were 0.2units/ml thrombin, 0.01mol/L CaCl2 and 0.7pmol/L tPA. Plasma samples were run in duplicate both with and without the addition of tPA. The absorbance was measured every 12 seconds for 90 minutes at 405nm.
Bio one microtitre plate
Manufactured by Greiner
Sourced in United States
The Bio-one microtitre plate is a laboratory equipment designed for a variety of assays and experiments. It provides a standardized platform for conducting multiple tests or analyses simultaneously in a compact format. The plate features a grid of small wells, allowing for the efficient use of sample materials and reagents.
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Lab products found in correlation
2 protocols using bio one microtitre plate
1
Murine Plasma Turbidity and Lysis Assay
2
Enzyme Kinetics of Purified Protease
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Enzyme kinetics Michaelis-Menten constant (Km), maximal velocity (Vmax), and kcat of the purified enzyme were determined using Z-Gly-Pro-pNA with substrate concentrations in the reaction mixture of 0.2, 0.5, 1, 1.5, 2 mM under the optimal assay conditions in the standard method, i.e. 60 o C and pH 6.6. An extinction coefficient for nitroaniline of 5.57 mM -1 cm -1 at 410 nm was used. The reactions were conducted by adding 20 µL of enzyme (containing 0.208 mg/ml protein) to buffer and substrate and the final volume was 250 µL in a microtitre plate well, which equated to a 1 cm path length. Absorbance was measured constantly by a BioTek Synergy 2 microplate Reader (USA) after substrate addition using a Greiner Bio-One micro-titre plate. The kinetic data were calculated from Lineweaver-Burk plots using the Michaelis -Menten equation.
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