Absorption Systems (Philadelphia, PA) performed the detection of the Pim1 inhibitor in the blood and brain of C57BL/6 mice injected with a dosage of 50 mg/kg. Standards were prepared in C57BL/6 mouse plasma containing sodium heparin as an anticoagulant, or in blank homogenized C57BL/6 mouse brain. The calibration curve was prepared to concentrations of 1000, 500, 250, 100, 50, 10, 5, and 2.5 ng/mL by serial dilution. Standard samples were treated identically to the study samples. Plasma and brain homogenate samples were extracted via acetonitrile precipitation on a Tomtec Quadra 96-Model 320 liquid handling system in a 96-well plate format. The procedure for sample extraction were as follows; (1) Add 55 μL of samples or standards into 2 mL polypropylene 96-well plate; (2) Using the Tomtec, add 50 μL of sample to 150 μL of acetonitrile (containing 100 ng/mL warfarin as an internal standard) that has been pre-loaded onto a Sirocco Protein Precipitation plate (Waters Corp.); (3) Using the Tomtec, mix the samples via air aspiration; (4) Apply vacuum and Cap for analysis.
Sirocco protein precipitation plate
Manufactured by Waters Corporation
The Sirocco Protein Precipitation plate is a laboratory equipment designed for the purpose of precipitating proteins from samples. It functions by providing a controlled environment for the precipitation process.
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Lab products found in correlation
2 protocols using sirocco protein precipitation plate
1
Quantification of Pim1 Inhibitor in Mice
2
Quantifying Plasma Citrulline and CRP in HIV Patients
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Four milliliters of venous blood was collected into EDTA-treated Vacutainer tubes (Becton Dickinson; Franklin lakes, NJ USA). Samples for complete blood counts were analyzed using a Coulter counter at the Uganda Cancer Institute laboratory. HIV sero-status was assessed at the MNU side laboratory using Determine HIV-1/2 (Abbot Laboratories USA) rapid tests and HIV-1/2 Stat-Pak Dipstick Assay kit for children aged ≥18 mo. For those <18 mo of age, their HIV status was confirmed using an HIV DNA/polymerase chain reaction (PCR) test done at Baylor HIV clinic. Plasma was obtained by centrifuging at 1300–2200 g at ambient temperature for 10 min, and stored at −80°C prior to shipment on dry ice to the University of Copenhagen, Denmark. Plasma C-reactive protein (CRP) was analyzed by the high-sensitivity kit on an ABK Pentra 400 (Horiba; Montpellier, France). Detection of plasma citrulline has been described in detail elsewhere (40 (link)). Briefly, the LC-MS method (41 ) was used. Samples were processed with Sirocco protein precipitation plate (Waters) and separated on an ACQUITY UPLC HSS T3 Column (Waters) using an ultra-performance LC in tandem with triple quadrupole detector MS. l -Citrulline-4,4,5,5-d4 (Sigma-Aldrich) was used as an internal standard. Quantification of citrulline was carried out using QuanLynx (Waters).
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