Overnight peripheral fasting blood samples were taken from all subjects, and the serum was either isolated and used for analyses, or stored at −80 °C. All samples were analyzed by specialized clinical laboratory medical personnel. The laboratory analyses included measurements of total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), triglycerides, glucose, creatinine (CRE), HbA1c(%), folic acid, vitamin B12, C-reactive protein (CRP), aspartate aminotransferase (ASP), alanine aminotransferase (ALT) and erythrocyte sedimentation rate (ESR). Serum levels of interleukine 6 (IL-6) were determined by enzyme-linked immunosorbent assay (ELISA), using the Diaclone Human IL-6 High Sensitivity ELISA kit (Diaclone SAS, Besancon Cedex, France). The estimated glomerular filtration rate (eGFR) was calculated by the modification of diet in renal disease (MDRD) and the chronic kidney disease epidemiology collaboration (CKD-EPI) equations [13 (link)]:
Human il 6 high sensitivity elisa kit
Manufactured by Diaclone
Sourced in France
The Human IL-6 High Sensitivity ELISA Kit is a quantitative in vitro assay designed for the measurement of human interleukin-6 (IL-6) in biological samples such as serum, plasma, and cell culture supernatants. The kit utilizes the enzyme-linked immunosorbent assay (ELISA) technique to detect and quantify IL-6 levels with high sensitivity.
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7 protocols using human il 6 high sensitivity elisa kit
1
Comprehensive Metabolic and Inflammatory Profiling
2
Comprehensive Metabolic Profiling of Participants
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Overnight peripheral fasting blood samples were taken from all subjects. The sera were isolated, used for analyses or stored at −80 °C. All samples were analyzed by specialist clinical laboratory personnel. The biochemical analyses included measurements of glucose, HbA1c (%), folate, vitamin B12, total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides, creatinine (CRE), C-reactive protein (CRP), 25(OH)D, aspartate aminotransferase (ASP), alanine aminotransferase (ALT) and erythrocyte sedimentation rate (ESR). Serum levels of interleukin 6 (IL-6) were determined with an enzyme-linked immunosorbent assay (ELISA), using the Diaclone Human IL-6 High Sensitivity ELISA kit (Diaclone SAS, Besancon Cedex, France). The laboratory staff was kept unaware of the genotypes of the subjects.
3
Preoperative Blood Biomarkers Analysis
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Blood samples were drawn in the morning after 12-h overnight fasting at the time of surgery (pre‐surgery) and during the follow‐up visit. Biochemical parameters measured included total cholesterol (TC), HDL-cholesterol (HDL-C), LDL-cholesterol (LDL-C), triglycerides (TG), fasting blood glucose (FBG), HbA1c (%), folic acid, vitamin B12, C-reactive protein (CRP), aspartate aminotransferase (ASP), alanine aminotransferase (ALT), insulin, and IL-6. Laboratory analyses were performed by routine laboratory methods. Serum levels of interleukin 6 (IL-6) was determined by the ELISA method using the Diaclone Human IL-6 High Sensitivity ELISA kit (950.035.192). Serum levels of insulin were determined by the ELISA method using the DRG® Insulin ELISA (EIA-2935) kits. Insulin resistance was assessed using the homeostasis model assessment [HOMA-IR index = (fasting glucose in mmol/L * fasting insulin in µIU/mL)/22.5] [31 (link)].
4
Lipid Profile and Inflammation in Psoriasis
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Blood samples were collected after 12 h to 14 h of fasting, and serum was obtained by low-speed centrifugation at 4°C. The concentrations of glucose, TC, high-density cholesterol (HDL-C), LDL-C, VLDL-C, and triglycerides (TG) were measured in fresh serum samples with standard diagnostic methods used in our hospital laboratory (Cobas Integra 400; commercially available reagents, Roche Diagnostics, Japan). Enzyme-linked immunosorbent assays (ELISA) were used to measure the serum concentration of IL-6 in frozen serum samples stored at –70°C until used (Human IL-6 High Sensitivity ELISA Kit, Diaclone, Besancon, France). We calculated the LDL : HDL ratio and the Casteli index according to the following formula: TC/HDL-C. Based on standard cholesterol cut-off, we classified patients into those with normal blood lipids (concentrations of TC < 200 mg/dl, LDL-C < 135 mg/dl, TG < 150 mg/dl: 35 patients with psoriasis; 13 patients with PsA) and abnormal blood lipids (concentrations of TC ≥ 200 mg/dl, LDL-C ≥ 135 mg/dl, TG ≥ 150 mg/dl: 27 patients with psoriasis; 18 patients with PsA).
5
Comprehensive Metabolic Profiling of Subjects
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Overnight peripheral fasting blood samples were taken from all subjects, and the serum was either isolated and used for analyses or stored at − 80 °C. All samples were analyzed by specialized clinical laboratory medical personnel. The laboratory analyses included measurements of total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides, glucose, HbA1c (%), 25(OH)D, C-reactive protein (CRP), 25(OH)D, ASP, ALT, and erythrocyte sedimentation rate (ESR). Serum levels of interleukin 6 (IL-6) was determined by the ELISA method using the Diaclone Human IL-6 High Sensitivity ELISA kit.
6
Quantification of NET Markers in Plasma
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Different markers of NETs were quantified in the plasma sample obtained before surgery, following the strategy addressed in previous studies (11 (link)–13 (link)), according to manufacturer´s instructions. Namely, cell-free DNA (cfDNA, Quant-iT PicoGreen dsDNA kit, Life Technologies, Eugene, OR, USA), nucleosomes (Cell Death Detection ELISAPLUS kit, Roche, Mannheim, Germany), calprotectin (Human Calprotectin ELISA kit, Hycult Biotech, Uden, The Netherlands), myeloperoxidase (MPO, Human MPO ELISA kit, Abnova, Taoyuan, Taiwan) and neutrophil elastase (NE, PMN Elastase Human ELISA Kit, Abcam, Cambridge, UK).
As inflammation marker, we quantified interleukin 6 (IL-6) (Human IL-6 High Sensitivity ELISA Kit, Diaclone, Besançon, France) following manufacturer´s instructions.
As inflammation marker, we quantified interleukin 6 (IL-6) (Human IL-6 High Sensitivity ELISA Kit, Diaclone, Besançon, France) following manufacturer´s instructions.
7
Genetic Variants in IL-6 and STAT3
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Genomic DNA was isolated from all the blood samples with Blood Mini A&A Biotechnology (A&A Biotechnology, Gdansk, Poland) according to the instructions of the manufacturer. Analysis of the polymorphic variants IL-6 rs1800795 (-174 G/C) and in the STAT3 gene: rs2293152 and rs4796793 were analyzed with the use of polymerase chain reaction with sequence-specific primers (SSP-PCR). Serum IL-6 levels were measured with an enzyme-linked immunoabsorbent assay (ELISA) standard kit (Human IL-6 High Sensitivity ELISA kit, Diaclone SAS, France).
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