All experiments were performed per the NIH Guide for the Care and Use of Laboratory Animals and were approved by Institutional Animal Care and Use Committees at Brigham Young University-Provo. Wild type C57BL/6 mice were sacrificed and were perfused ventricularly with sodium heparin solution in PBS (H3393–100KU, Sigma-Aldrich, MO, USA). The lungs were then excised and fixed in 4% PFA overnight at 4 °C. The porcine lungs were gifted by Circle V Meats Co. (UT, USA). The lungs were received and processed within the hour of sacrifice. Lung sections were washed thrice with heparinized PBS for 10 min each and visible blood clots were removed. The lungs were then sectioned into 1-inch cubes and fixed overnight with 4% PFA at 4 °C.
H3393 100ku
Manufactured by Merck Group
Sourced in United States
H3393–100KU is a laboratory product manufactured by Merck Group. It is a specialized piece of equipment designed for use in research and scientific applications. The core function of this product is to provide a controlled environment for various experimental and analytical processes. Further details on the intended use or specific applications of this product are not available.
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Lab products found in correlation
2 protocols using h3393 100ku
1
Murine and Porcine Lung Perfusion and Fixation
2
In Vivo BBB Permeability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Blood-brain barrier (BBB) permeability was examined with the in vivo permeability assay as described previously (17) (link). Briefly, FITC-dextran (70 kDa, Sigma catalog no. 46945; 1 mM in 50 μL sterile PBS per injection) was injected intracardially into mice with or without NEC. After 5 min, mice were perfused transcardially with 10 mL of heparinized PBS (Sigma, H3393-100KU; 10 U/mL) by slow injection. After perfusion, brains were collected, weighed, homogenized in 200 μL of PBS, and centrifuged at 16,000 g for 20 min at 4°C. The sample supernatant (20 μL) was collected, mixed with 100 µL of PBS, and further loaded into black 96-well plates (Corning, CLS3925). Measurement of relative fluorescence units (RFUs) was obtained with a 96-well plate reader (Spectramax; Molecular Devices) at 490/520 nm. Each sample was measured in triplicate, and the average value was used as one biological data point.
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