Circulating CTRP7 levels were determined with an ELISA kit (sk00396-09, Aviscerabio science Inc., MA. USA) according to the manufacturer’s protocol. The detection limit of serum CTRP7 levels was 5-320 μg/L, and the intra- and inter-assay coefficients of variation (CV) were less than 5% and 10%, respectively. The ELISA kit has high sensitivity, good specificity for human CTRP7 detection without obvious cross-reaction, and interference. Circulating APN levels were also measured with an ELISA kit following the manufacturer’s protocol (Aviscera Bioscience, sk00010-02). Intra- and inter-assay CV were 8% and 10%, respectively.
ELISA kit
Manufactured by Aviscera Bioscience
Sourced in United States
The ELISA kit is a laboratory tool used for the detection and quantification of specific proteins or other biomolecules in a sample. It is a sensitive immunoassay technique that utilizes antibodies to capture and detect the target analyte. The kit includes all the necessary reagents and materials required to perform the assay.
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Lab products found in correlation
ELISA kits, R&D Systems (3 mentions)
ELISA kits, RayBiotech (2 mentions)
ELISA kits, Elabscience (2 mentions)
Hitachi-912 Autoanalyser, Hitachi (1 mentions)
ELISA kit, Echelon Biosciences (1 mentions)
ELISA kits, Thermo Fisher Scientific (1 mentions)
ELISA kit, Monobind (1 mentions)
Enzyme-linked immunosorbent assay (ELISA), Bio-Rad (1 mentions)
Sk00010-01, Aviscera Bioscience (1 mentions)
Synergy H1TM, Agilent (1 mentions)
Microplate absorbance reader, Bio-Rad (1 mentions)
DRP300, R&D Systems (1 mentions)
ELISA kit, Adipogen (1 mentions)
27 protocols using ELISA kit
1
Serum CTRP7 and APN Quantification
2
Lipid and Inflammation Biomarker Changes
Twenty-four hours before the first training session and 48 h after the protocol, fasting blood samples were drawn. Serum samples were centrifuged for 15 min (3,000 rpm, 4 °C). Serum sICAM-1 concentrations were determined using the Soluble ICAM-1/CD54 (Human) ELISA Kit (EIA-SK00250-02, Aviscera Bioscience, Santa Clara, CA, USA) with the sensitivity of 4.88 pg/ml. Serum IL-6 concentrations were determined using the ELISA Kit (EA550799-02, Aviscera Bioscience) with the sensitivity of 0.5 pg/ml. The HDL-C, LDL-C, TG, TC, and hs-CRP were assessed using the Pars Azmoon Company kits (Tehran, Iran) with help of the automatic analysis machine, Prestige 24i.
3
Quantification of Soluble RAGE Using ELISA
Human soluble receptor for advanced glycosylation end products (sRAGE) ELISA kit (Aviscera Bioscience) was used to quantify the amount of secreted sRAGE. The samples and standards were loaded in a pre-coated 96-well microplate and incubated for 2 hours at room temperature. After washing the wells four times with washing buffer, 100 μl of detection antibody was added to each well and incubated at room temperature. After 2 hours of incubation, the wells were rinsed for four times with washing buffer and then 100 μl of ARIG-HRP conjugated working solution was added to each well incubated at room temperature for 1hour at room temperature. After washing the plate with washing buffer, 100 μl of Substrate solution to each well followed by incubating for 10 minutes at room temperature. Finally, 100 μl of Stop solution added each well and optical density was determined at 450 nm.
4
Plasma PTX3 Measurement and Genomic DNA Extraction
We drew 3 ml of venous blood from every subject on an empty stomach in the morning; 1 ml of the blood sample was centrifuged to separate plasma at 4000 rpm for 10 min. Plasma PTX3 was measured using an enzyme-linked immunosorbent assay (ELISA) kit (Aviscera Bioscience Inc., USA) according to the manufacturer’s instructions. The other 2 ml of the blood sample was used to extract genomic DNA by use of the TIANGEN blood genomic DNA extraction Kit (BEIJING TIANGEN BIOTECH CO., LTD) and stored at –20°C for genotyping in the next step.
7
Blood Lipid and Glucose Profiling
Blood samples were taken from all subjects after they had fasted for nearly 12 hours. In the next step, serum samples were separated by centrifugation at 2,800×g for 10 min. Subsequently, the lipid profile including total cholesterol, triacylglycerol (TG), high-density lipoprotein cholesterol (HDL-C), and fasting blood sugar (FBS) was assayed by use of a Hitachi-912 Autoanalyser. Low-density lipoprotein cholesterol (LDL-C) levels were measured according to the Friedewald formula. Glycated hemoglobin (HbA1c) was determined by using a BioSystems kit according to the manufacturer's protocol. The concentration of serum betatrophin was measured by using an ELISA kit (Aviscera Bioscience). The ELISA was carried out according to the manufacturer's protocol. All samples were tested in duplicate, and samples with a coefficient of variation (CV) greater than 15% were excluded. Insulin concentration was determined by use of an immunoassay kit (Monobind Inc) and resistance to insulin was evaluated according to the homeostatic model assessment (HOMA-IR) formula [insulin (µU/mL)×glucose (mmol/L)/22.5].15 (link)
8
Adiponectin and CTRP15 Measurement
Circulating levels of adiponectin were measured using the ELISA technique (previously described) [36 ]. The serum levels of CTRP15 were detected using the ELISA kit (Aviscera Bioscience, Inc., USA; catalog number: SK00393-15). Following the manufacturer’s instructions, we used the intra- and inter-assay coefficients of variants (CV) of 6% and 10%, respectively.
9
BDNF Quantification in Animal Sera
Three batches of animals (MSA, n= 8 and SSA, n=8 per batch) were used for trunk blood collection on self-administration days 3, 5, and 7, respectively. Blood BDNF levels were assayed using the ELISA kit (Aviscera Bioscience, Santa Clara, CA). Serum samples were obtained from trunk blood (2 hours after the session) and were centrifuged at 2000 rpm for 20 minutes. Samples were diluted to 1/40 with dilution buffer, and then the plate was incubated for 2 hours with gentle shaking. The wells were washed 4 times and 100 μL of antibody working solution was added. After a second incubation on the shaker for 2 hours, the wells were washed 4 times. Then 100 μL of conjugate working solution was added followed by a 3rd incubation for 1 hour. After washing the wells 4 more times, 100 μL of substrate solution was added and the wells were incubated for 8 minutes on a shaker. A stop solution of 100 μL was added, and after incubation, the optical density was detected at 450 nm using an Infinite M200 Pro Microplate Reader (Tecan US, Morrisville, NC). To test for the possibility of intra-assay variation, each sample was tested in duplicate. This assay protocol followed the manufacturer’s instructions and a previous study (Polacchini et al., 2015 (link)).
10
Plasma Myonectin Quantification via ELISA
Plasma myonectin levels were measured using a commercially available ELISA kit (AVISCERA BIOSCIENCE, INC. CA, USA; Catalog number # SK00393-10). Plasma samples were diluted 4 times with sample dilution buffer for myonectin measurement as per manufacturer’s recommended protocol. This assay has a coefficient of variation of <12% and minimum detection limit of 1 ng/ml.
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