For ELISA, serum CD62P (cat. no. MPS00; R&D Systems, Inc.) and VEGF-a (cat. no. MMV00; R&D Systems, Inc.) were quantified using commercially available anti-mouse ELISA kits, according to the manufacturer's instructions. All assays were run in duplicate at a suitable dilution. The absorbance was read by a microplate reader (Multiskan GO, Thermo Fisher Scientific, Inc.) set at 450 nm and corrected at 540 nm.
Mps00
Manufactured by R&D Systems
Sourced in United States, United Kingdom
The MPS00 is a laboratory equipment product. It performs core functions related to the intended application of the device. No additional details can be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
4 protocols using mps00
1
Mouse Serum Biomarker ELISA Quantification
2
Plasma DNA and Coagulation Factors Analysis
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Blood was collected into 1/10 final volume of 3.2% (wt/vol) sodium citrate anticoagulant. After centrifugation of whole blood at 3300 × g for 5 min, plasma was collected and centrifuged at 16100 × g for 5 min to remove remaining cellular components. Plasma DNA was measured using the Quant-iT PicoGreen dsDNA Assay Kit (Thermo Fisher Scientific, P7589). TAT complexes (Abcam, ab137994) and soluble P-selectin (R&D systems, MPS00) were measured by ELISA according to the manufacturer’s instructions.
3
Quantification of Soluble E-Selectin and IL-6
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Soluble E-selectin (sEsel), and IL-6 enzyme-linked immunosorbent assays (ELISA) were performed according to the manufacturer’s instructions (R&D Systems, Inc.; Minneapolis MN, USA; Cat#: MES00 & MPS00), n=3-5. Blood for ELISA was collected via cardiac puncture at the time of sacrifice and plasma prepared by centrifugation at 8,500 rpm for 10 min.
4
Quantifying Hemostasis Biomarkers in Plasma
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Blood was collected via intracardiac puncture and immediately mixed with ethylenediaminetetraacetic acid (EDTA). The EDTA-blood solution was centrifuged for 15 min at 3000g, as previously described [9] (link). Plasma was collected and snap-frozen in liquid nitrogen. Levels of proteins of interest were measured using colorimetric enzyme-linked immunosorbent assays species-specific following the manufacturer's instruction. Specifically, DY3178 was used for TF (R&D systems, Minneapolis, MN, USA), ab233615 was used for tissue-plasminogen activators (tPA; Abcam, Cambridge, UK), DY3828 was used for plasminogen activator inhibitor 1 (R&D systems, Minneapolis, MN, USA), abx258705 was used for D-dimer (Abbexa, Cambridge, UK), MPS00 was used for soluble P-selectin (R&D systems, Minneapolis, MN, USA), NBP2-68171 was used for von Willebrand factor (vWF; Novus Biologicals, Cambridge, UK), NBP2-60632 was used for antithrombin III (Novus Biologicals, Cambridge, UK), and ab137994 was used for thrombin-antithrombin complexes (Abcam, Cambridge, UK).
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