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Human tnf α quantikine hs elisa

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The Human TNF-α Quantikine HS ELISA is a highly sensitive enzyme-linked immunosorbent assay (ELISA) designed to quantitatively measure human tumor necrosis factor alpha (TNF-α) levels in cell culture supernates, serum, and plasma.

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3 protocols using human tnf α quantikine hs elisa

1

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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2

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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3

Metabolic and Adipokine Profiles

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Fasting blood samples were collected on the morning of surgery after a 12h-overnight fast. From these samples, cholesterol and triglyceride levels in both plasma and lipoprotein fractions were measured (21) .
The homeostatic model assessment of insulin resistance (HOMA-IR) was calculated from fasting glucose and insulin levels as previously described (22) . Enzyme-linked immunosorbent assay was performed on these samples for the following adipokines: leptin (Human Leptin ELISA kit; EMD Millipore; Billerica, MA, USA), adiponectin (Human Adiponectin ELISA Kit; B-Bridge International Inc; Santa Clara, CA, USA), interleukin-6 (IL-6) (Human IL-6 Quantikine HS ELISA; R&D Systems; Minneapolis, MN, USA) and tumor necrosis factor-α (TNF-α) (Human TNF-α Quantikine HS ELISA; R&D Systems; Minneapolis, MN, USA).
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