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4 protocols using human icam 1 cd54 allele specific quantikine elisa kit

1

Serum Biomarkers in ELISA Analysis

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The serum levels of ICAM-1, VCAM-1, E-selectin, and S100 protein were measured using the enzyme-linked immunosorbent assay (ELISA) in duplicate. The Human ICAM-1/CD54 Allele-specific Quantikine ELISA Kit (DCD540; R&D Systems, Minneapolis, MN, USA), Human sVCAM-1/CD106 Quantikine ELISA Kit (DVC00, R&D Systems), Human sE-Selectin/CD62E Quantikine ELISA Kit (DSLE00, R&D Systems), and Human S100 ELISA Kit (MBS2503148; MyBioSource Inc., San Diego, CA, USA) were used, respectively.
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2

Biomarker Measurement Protocol for Plasma

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Blood extraction was performed between 8:00 and 9:00 a.m. following an overnight fast by nurses in the Primary Health Care Centers. All participants were instructed to not exercise 8 h before the blood test. Blood samples were obtained from the antecubital vein and collected in EDTA tubes for plasma analyses. Tubes were immediately transferred to the IGTP-HUGTP Biobank integrated in the Spanish National Biobanks Network of Instituto de Salud Carlos II (PT13/0010/0009) and Tumor Bank Network of Catalonia, and they were processed following standard operating procedures with the appropriate approval of the Ethical and Scientific Committees. Plasma aliquots were stored at −80°C.
As Level 1 outcomes, we obtained peripheral BDNF levels using an ELISA kit (Human Free BDNF Quantikine ELISA Kit; R&D Systems, Minnesota, USA). The rest of the molecular markers were selected according to the Projecte Moviment trial (Castells-Sánchez et al., 2019 (link)). TNF-α, ICAM-1, HGF, and SDF1-α levels were analyzed quantitatively using the corresponding ELISA immunoassay method (Human TNF-α Quantikine HS ELISA, Human ICAM-1/CD54 Allele-specific Quantikine ELISA Kit, Human HGF Quantikine ELISA Kit, Human CXCL12/SDF-1 alpha Quantikine ELISA Kit; R&D Systems, Minnesota, USA).
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3

Quantifying Plasma Biomarkers in Healthy Adults

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Nurses in the Primary Health Care Centers obtained blood samples from the antecubital vein in EDTA tubes for plasma analyses between 8:00 and 9:00 a.m. All participants were instructed to fast overnight and not exercise 8 h before the blood test. Tubes were immediately transferred to the IGTP-HUGTP Biobank integrated in the Spanish National Biobanks Network of Instituto de Salud Carlos II (PT13/0010/0009) and Tumor Bank Network of Catalonia. They were processed following standard operating procedures with the appropriate approval of the Ethical and Scientific Committees and plasma aliquots were stored at −80°C.
We obtained peripheral BDNF levels using an ELISA kit (Human Free BDNF Quantikine ELISA Kit; R&D Systems, Minnesota, United States). The Proteome Profiler Human™ XL Cytokine Array was used to semi-quantitatively analyze a panel of 105 targeted cytokines (R&D Systems, MN, United States). Based on the results of the array and previously cited literature, TNF-α, ICAM-1, HGF, SDF1-α levels were selected to be quantitatively analyzed using the corresponding ELISA immunoassay method (Human TNF-α Quantikine HS ELISA, Human ICAM-1/CD54 Allele-specific Quantikine ELISA Kit, Human HGF Quantikine ELISA Kit, Human CXCL12/SDF-1 alpha Quantikine ELISA Kit; R&D Systems, Minnesota, United States).
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4

Measuring Soluble Adhesion Molecules

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Soluble ICAM-1 and VCAM-1 (sICAM-1 and sVCAM-1, respectively) levels were determined in maternal total serum from MPH and MSPH women by ELISA kits (Human ICAM-1/CD54 Allele-specific Quantikine ELISA Kit and Human VCAM-1/CD106 Quantikine ELISA Kit, respectively; R&D Systems, Minneapolis, MN, USA), as described [25 (link)]. The results were expressed as ng/mL, according to calibration curves.
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