β-TCP scaffolds seeded with JPCs were cultivated in vitro for 15 days. The scaffolds were then transferred into 12 mL polypropylene round-bottom tubes (Becton Dickinson, Franklin Lakes, NJ, USA) containing 11 mL blood. Control tubes contained the same amount of fresh blood without scaffolds. The incubation with blood was performed at 37 °C for 90 min using a tube rotator (neoLab, Heidelberg, Germany) with 10 rpm. After 90 min, blood was transferred into monovettes containing ethylenediaminetetraacetic acid (EDTA) (Sarstedt Inc., Nümbrecht, Germany) for the analysis of complement activation and detection of cell numbers. Blood cell numbers were determined using a Micros 60 cell counter (ABX Diagnostics, Montpellier, France). A citrate solution containing monovettes (Sarstedt Inc.) was used to analyze PMN-elastase and TAT. To analyze β-TG concentrations, blood was transferred to citrate-theophylline-adenosine-dipyridamole (CTAD) containing monovettes (BD Biosciences Inc.) and stored for 15 min on ice. EDTA and CTAD monovettes were centrifuged at 2500× g for 20 min at 4 °C. Citrate blood monovettes were centrifuged at 1800× g for 18 min at RT. The plasma of each sample was shock frozen in liquid nitrogen and stored at −20 °C (EDTA and citrate plasma) or −80 °C (CTAD plasma) until further investigations.
Monovettes
Manufactured by Sarstedt
Sourced in Germany
Monovettes are closed blood collection systems designed to collect and transport blood samples. They are made of high-quality plastic and feature a secure closure to prevent leakage. Monovettes are available in a range of sizes and with various additives to accommodate different blood testing requirements.
Automatically generated - may contain errors
Lab products found in correlation
Ethylene diamine tetraacetic acid (edta), by Sarstedt (2 mentions)
Cobas 6000 system, by Roche (1 mentions)
Sorvall rc6 centrifuge, by Thermo Fisher Scientific (1 mentions)
Dimension clinical chemistry system, by Siemens (1 mentions)
Scgm media, by CellGenix (1 mentions)
Dnase, by Thermo Fisher Scientific (1 mentions)
Rnalater, by Thermo Fisher Scientific (1 mentions)
Cellometer x2, by Revvity (1 mentions)
6 well plate, by Corning (1 mentions)
Edta monovette, by Sarstedt (1 mentions)
Oasis max spe system plates, by Waters Corporation (1 mentions)
Quattro premier xe triple quadrupole mass spectrometer, by Waters Corporation (1 mentions)
Labofuge 400r, by Thermo Fisher Scientific (1 mentions)
Pentra 400, by Horiba (1 mentions)
Enzyme linked immunosorbent assay, by Mercodia (1 mentions)
Enzymatic colorimetric assay, by Roche (1 mentions)
Cobas c analyzer, by Roche (1 mentions)
Ultrapure dnase rnase free distilled water, by Thermo Fisher Scientific (1 mentions)
Qiaamp circulating nucleic acid kit, by Qiagen (1 mentions)
Ez dna methylation kit, by Zymo Research (1 mentions)
21 protocols using monovettes
1
In Vitro Blood Compatibility of β-TCP Scaffolds
2
Resveratrol Genotoxicity Evaluation
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The experimental group was composed of healthy, non-smoking female volunteers with similar nutritional conditions. Participants received one capsule of 50 mg RSV daily under fasting conditions for 15 consecutive days. A sample of about 5 mL peripheral blood was drawn in labelled tubes containing lithium heparin as anticoagulant (Monovettes®; Sarstedt, Nümbrecht, Germany) at the beginning and end of this treatment. Immediately afterwards, two aliquots of blood (1 mL each) were separated and centrifuged for 5 min at 1000 rpm at 20 °C. Plasma was removed and replaced for an equal volume of RPMI, and the pellets were resuspended. One aliquot was irradiated with 4 Gy and the other was used as experimental control. Genotoxic damage was assessed with the alkaline comet assay.
3
Biochemical Markers in Fasting Samples
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Blood samples were collected in the fasting state. Liver enzymes (aspartate transaminase [AST], alanine transaminase [ALT], 𝛾-glutamyltransferase [𝛾-GT]), creatinine, uric acid, bilirubin (total), and thyroid-stimulating hormone (TSH) were analyzed in serum by SynLab (Munich, Germany). Additionally, plasma (EDTA KE monovettes, Nümbrecht, Sarstedt) was collected and centrifuged at 1800 × g for 10 min at 4 °C. Serum (Sarstedt monovettes) was collected, left for 20 min to allow clotting, and was finally centrifuged (2500 × g for 10 min at 4 °C). Plasma and serum were aliquoted and stored at -80 °C for later measurement of selected biochemical parameters.
4
Plasma Biomarker Quantification Protocol
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Venous blood was collected into 0.3 mL citrate buffer tubes (Monovettes, Sarstedt, Nuremberg, Germany). Plasma concentrations of fibrinogen (lower limit of detection, LOD: 30 mg/dL) and VWF antigen (lower LOD: 270 mg/L) were determined with a multiplexed particle-based flow cytometric assay (Cytolab, Regensdorf, Switzerland). Plasma D-dimer concentration (lower LOD: 0 ng/mL) was determined with an enzyme-linked immunosorbent assay (Technozym D-dimer, Technoclone, Vienna, Austria). Inter-and intra-assay coefficients of variation were <15% for all analyses. CRP plasma levels were determined with the cobas 6000 system (Roche Diagnostics, Switzerland).
5
Biomarker Measurement Protocol for Blood Samples
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Blood samples were drawn by experienced nurses using EDTA-coated vacutainers (monovettes®, Sarstedt GmbH, Nümbrecht, Germany). Hb was measured using an automated Counter (ACT 5 diff counter, Beckman Coulter International S.A., Nyon, Switzerland). A control material of 3 levels (Coulter AC.T5 diff Control Plus) was analyzed with each set of measurements of Hb. Plasma was separated for ferritin and CRP measures by centrifugation (Sorvall RC6 + centrifuge, ThermoFisher Scientific, Osterode, Germany). Ferritin and CRP were measured with the Siemens Dimension® Clinical Chemistry system using Flex® reagent cartridges. Ferritin was measured using the enzyme immunoassay method and CRP using the C-Reactive Protein Extended Range (RCRP) method based on a particle enhanced turbidimetric immunoassay (PETIA) technique. Two level serum control material (Liquid Assayed Multiqual Premium) were analyzed with each ferritin and CRP measurements.
6
Confrontation Assay for Neutrophil-Candida Interaction
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Venous blood of healthy volunteers was collected in ethylenediaminetetraacetic acid monovettes (Sarstedt). Polymorphonuclears (PMNs) were subsequently purified as described (67 (link),68 (link)). Immune cells were suspended in SCGM media (CellGenix), counted with a cellometer X2 (Nexcelom) and immediately subject to confrontation assay.
Confrontation of 3 × 106C. glabrata cells with 6 × 106 PMN was performed in one well of a 6-well plate (Corning) in a total volume of 3-ml SCGM media. Following the incubation period, 20-U DNase (Invitrogen) were added to the co-incubation to dissolve DNA based structures, which trapped fungal material. Co-incubations were flooded with five volumes of RNAlater (Ambion). The solution was mixed with an equal volume of 4°C H2O to lyse neutrophils. Fungal cells were harvested by centrifugation at maximum g for 5 min and subject to immediate RNA isolation.
Confrontation of 3 × 106C. glabrata cells with 6 × 106 PMN was performed in one well of a 6-well plate (Corning) in a total volume of 3-ml SCGM media. Following the incubation period, 20-U DNase (Invitrogen) were added to the co-incubation to dissolve DNA based structures, which trapped fungal material. Co-incubations were flooded with five volumes of RNAlater (Ambion). The solution was mixed with an equal volume of 4°C H2O to lyse neutrophils. Fungal cells were harvested by centrifugation at maximum g for 5 min and subject to immediate RNA isolation.
7
Trauma-Induced Blood and Fracture Hematoma Analysis
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Full blood samples were drawn exactly before induction of trauma (0 h), after trauma and resuscitation (2.5 h), and during observation period at 14 h, 24 h, and 48 h using monovettes (SARSTEDT AG & Co, Germany). Fracture hematoma was extracted under sterile conditions by puncturing the fracture zone. Hematoma was collected in an EDTA monovette (SARSTEDT AG & Co, Germany). After centrifugation, serum was removed and stored at −80°C for further analysis. After an observation period of 48 hours the animals were sacrificed. Data was collected using a Filemaker Database (FilmakerPro5.0, Filemaker Inc.); access was limited by password protection. Security setup was done every 48 h.
8
Serum Testosterone and Cortisol Determination
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Blood samples were obtained from an antecubital vein in monovettes (Sarstedt, Germany) in the early morning before 8 am after an overnight fast and transferred within 1 h after sampling to the laboratory of the University Hospital Essen for analyses. Serum and plasma aliquots were stored at −80°C until testosterone and cortisol were determined by liquid-chromatography tandem mass spectrometry [LC-MS/MS; (23 (link), 24 (link))]. In brief, the stored sample aliquots, aliquots of calibrator and controls with a volume of 0.1 mL were combined with the internal standard mixture to monitor recovery. All samples were extracted using Oasis MAX SPE system Plates (Waters, Milford, MA, USA). LC-MS/MS was performed using a Waters Quattro Premier/Xe triple-quadrupole mass spectrometer connected to a Waters Acquity (Waters, Milford, MA, USA; Table 1 for details on assays).
To consider the biologically active fraction of testosterone and for reasons of comparability with previous studies, free testosterone (FT) levels were calculated according to Vermeulen et al. (25 (link)).
To consider the biologically active fraction of testosterone and for reasons of comparability with previous studies, free testosterone (FT) levels were calculated according to Vermeulen et al. (25 (link)).
9
Plasma Biomarker Measurement Protocol
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Venous blood samples were collected into precooled potassium ethylenediaminetetraacetic acid (EDTA) Monovettes (Sarstedt, Leicester, UK) and centrifuged at 1165g for 10 min at 4°C (Labofuge 400R; Thermo Scientific, Langenselbold, Germany). The plasma supernatant was aliquoted and stored at -80°C. Plasma concentrations of TAG, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, glucose, and high-sensitivity C-reactive protein were measured in duplicate using a benchtop analyzer (Pentra 400; HORIBA Medical, Montpellier, France), whereas insulin concentrations were measured in duplicate using an enzyme-linked immunosorbent assay (Mercodia, Uppsala, Sweden). The within-batch coefficient of variation for each assay was as follows: 1.0% for TAG, 0.4% for total cholesterol, 0.6% for highdensity lipoprotein cholesterol, 0.6% for low-density lipoprotein cholesterol, 0.4% for glucose, 3.3% for insulin, and 1.6% for high-sensitivity C-reactive protein.
10
Automated Lipid Quantification in Plasma
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TC and HDL-C were measured from heparin-coated Monovettes (Sarstedt Monovette orange). Analyses were performed using in vitro assays (enzymatic colorimetric assays, Roche, Mannheim, Germany) for the quantitative determination of blood lipids in human plasma on a Roche/Hitachi Cobas C Analyzer (Roche, Mannheim, Germany). The inter-and intra-assay CVs were ≤ 1.2% and ≤ 2.5%, respectively.
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