Plasma glucose was measured on a chemistry AutoAnalyzer by the glucokinase method. Insulin measurements were performed by a polyethylene glycol–accelerated double antibody radioimmunoassay method. hs-CRP and fibrinogen levels in plasma were measured immunochemically using Dade Behring reagent on the Behring Nephelometer II analyzer (BN II). tPA levels were measured in citrated plasma using an ELISA assay (Asserachrom tPA; Diagnostica Stago).
Asserachrom tpa
Manufactured by Diagnostica Stago
Sourced in France
The Asserachrom tPA is a laboratory equipment product manufactured by Diagnostica Stago. It is used to measure the levels of tissue plasminogen activator (tPA) in biological samples. tPA is an enzyme that plays a crucial role in the process of fibrinolysis, the breakdown of blood clots. The Asserachrom tPA device provides a quantitative analysis of tPA concentration, which can be useful in various clinical and research applications.
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5 protocols using asserachrom tpa
1
Comprehensive Metabolic Profiling Protocol
2
Quantification of Fibrinolytic Markers in Plasma
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Blood was collected in two plastic tubes containing 3.2% sodium citrate (BD Vacutainer®, Becton Dickinson, Franklin Lakes, NJ, USA) and 25 µg/mL hirudin (Dynabyte, Munich, Germany) at selected time points on the third day of hospitalization. After centrifugation (at 2500× g at +4 °C for 20 min), plasma was stored at –80 °C until further use in assays.
t-PA concentrations in human plasma samples were measured using ASSERACHROM® tPA (Diagnostica Stago, Asnières-sur-Seine, France). The detection limit of this assay was 1.5 ng/mL. The intra-assay coefficient of variation (CV) was between 5.1 and 7.3% and the inter-assay CV between 3.6 and 4.2%. PAI-1 concentrations were assessed using enzyme-linked immunosorbent assay kits (IMUBIND® Plasma PAI-1 ELISA, American Diagnostica Inc., Stamford, CT, USA). The lower detection limit for PAI-1 was 2.2 ng/mL. According to the manufacturer’s instructions, intra- and inter-assay CVs for PAI-1 were 5.4–6.6% and 6.9–9.0%, respectively. PAP complexes in human plasma were measured using PAP micro-ELISAs (DRG Instruments GmbH, Marburg, Germany). The analytical sensitivity of the DRG ELISA was 10 µg/mL. The activity of α2-AP was determined using a coagulometer Coag-Chrom 3003 (BioKsel, Grudziadz, Poland) and Bio-Ksel plasma kits. Normal values for α2-AP activity were 80–120%.
t-PA concentrations in human plasma samples were measured using ASSERACHROM® tPA (Diagnostica Stago, Asnières-sur-Seine, France). The detection limit of this assay was 1.5 ng/mL. The intra-assay coefficient of variation (CV) was between 5.1 and 7.3% and the inter-assay CV between 3.6 and 4.2%. PAI-1 concentrations were assessed using enzyme-linked immunosorbent assay kits (IMUBIND® Plasma PAI-1 ELISA, American Diagnostica Inc., Stamford, CT, USA). The lower detection limit for PAI-1 was 2.2 ng/mL. According to the manufacturer’s instructions, intra- and inter-assay CVs for PAI-1 were 5.4–6.6% and 6.9–9.0%, respectively. PAP complexes in human plasma were measured using PAP micro-ELISAs (DRG Instruments GmbH, Marburg, Germany). The analytical sensitivity of the DRG ELISA was 10 µg/mL. The activity of α2-AP was determined using a coagulometer Coag-Chrom 3003 (BioKsel, Grudziadz, Poland) and Bio-Ksel plasma kits. Normal values for α2-AP activity were 80–120%.
3
Mesothelial Cell Response to Thrombin
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Human mesothelial cells were incubated in 2 units of thrombin (Stago, - Gennevilliers, France) that had been added to serum-free conditioned HMC culture medium. To discard the thrombin from culture medium, twelve hrs later, the culture medium has been changed and cells were incubated again twenty-four hours with the serum-free conditioned HMC culture medium. The culture medium was collected for protein measurement. Incubated cells without thrombin addition to the culture medium were considered the control. Urokinase plasminogen activator (uPA) and plasminogen activator inhibitor (PAI-1) evaluations were performed using ELISA kits (Sigma-Aldrich, Saint-Quentin Fallavier, France), Tissue plasminogen activator (t-PA) evaluation was performed using Asserachrom® t-PA (Diagnostica Stago, Asnieres, France).
4
Coagulation Biomarkers Assessment Protocol
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Plasminogen activity in plasma was measured by a colorimetric assay (Stachrom Plasminogen, Diagnostica Stago, France). An enzyme-linked immunosorbent assay technique was applied to measure t-PA antigen level (Asserachrom t-PA, Diagnostica Stago, France). The activity of PAI-1 and α2-antiplasmin were determined by a colorimetric method (Stachrom PAI, Diagnostica Stago, France; Stachrom Antiplasmin, Diagnostica Stago, France). Quantitative measurement of vitronectin level was performed by an enzyme-linked immunosorbent assay (Imubind vitronectin, Americam Diagnostica GmbH). D-Dimer level was studied with an enzyme-linked immunosorbent technique (Imuclone D-dimer, American Daignostica GmbH).
The intra-assay coefficient of variation was <6%, except for PAI-1 activity (7.9%). The interassay coefficient of variation was <6%, except for PAI-1 activity (6.6%).
The intra-assay coefficient of variation was <6%, except for PAI-1 activity (7.9%). The interassay coefficient of variation was <6%, except for PAI-1 activity (6.6%).
5
Adiponectin, Inflammatory Markers Assay
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