Saliva samples were taken from each participant at 07:00, 07:30, and 07:45 every morning via sterile synthetic oral swabs (Sarstedt Salivette Cortisol, code blue). Each sampling period was 3 min. No food or fluids other than water were permitted until the final sample had been taken. The samples were refrigerated immediately and transferred to frozen storage (−80°C) within 3 h. Samples were thawed on the day of analysis and centrifuged at 1,500 ×g for 15 min. Cortisol concentrations were extracted in duplicate using an enzyme-linked immunosorbent assay (ELISA) technique (coefficient of variation 0.0–12.95%, mean 2.88%) designed for analysis of salivary cortisol (Salimetrics Salivary Cortisol Enzyme Immunoassay Kit).
Salivary cortisol enzyme immunoassay kit
Manufactured by Salimetrics
Sourced in United Kingdom, United States
The Salivary Cortisol Enzyme Immunoassay Kit is a laboratory equipment product designed to measure cortisol levels in saliva samples. The kit utilizes an enzyme-linked immunosorbent assay (ELISA) technique to quantify the concentration of cortisol, a steroid hormone, in the provided saliva samples.
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Lab products found in correlation
19 protocols using salivary cortisol enzyme immunoassay kit
1
Salivary Cortisol Sampling and Analysis
2
Saliva Cortisol Measurement Protocol
3
Cortisol Extraction and Quantification in Embryos
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The treated embryos were harvested at 5 dpf for cortisol ELISA assays. The cortisol extraction was performed following the protocol described in [50 (link)] with modifications. A group of 30 treated embryos pooled as one sample for ELISA was immobilized by ice-cold water and frozen in liquid nitrogen, and 3 ELISA samples were collected from each respective treatment. The samples were homogenized using a pellet mixer (Dr. Owl) and added with ethyl acetate for the collection of supernatant. The supernatant was evaporized using nitrogen gas, and lipid-containing extracted samples were dissolved in 60 μL of 0.2% BSA in PBS.
For the determination of whole-embryo cortisol levels, a cortisol ELISA kit (Salivary Cortisol Enzyme Immunoassay Kit, Salimetrics) was used according to the manufacturer’s instructions. Cortisol concentration values were obtained by performing 4 parameter logistic regression of the absorbance readings against a standard curve on a plate reader (TECAN Sunrise).
For the determination of whole-embryo cortisol levels, a cortisol ELISA kit (Salivary Cortisol Enzyme Immunoassay Kit, Salimetrics) was used according to the manufacturer’s instructions. Cortisol concentration values were obtained by performing 4 parameter logistic regression of the absorbance readings against a standard curve on a plate reader (TECAN Sunrise).
4
Salivary Biomarkers Measurement Protocol
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Participants used saliva collection aids (SCA; Salimetrics, State College, PA, USA) to collect their saliva into collection vials. Salivary samples were stored at 20 °C until analysis by immunoassay kits (Salimetrics) for salivary cortisol and s-IgA levels. Salivary cortisol and s-IgA levels were determined using an enzyme-linked immunosorbent assay with the following kits, respectively: salivary cortisol enzyme immunoassay kit and secretory immunoglobulin A salivary immunoassay kit (Salimetrics LLC, Carlsbad, CA, USA) [34 (link)].
5
Saliva Cortisol Enzyme Immunoassay
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The samples were analyzed using a commercial enzyme immunoassay method (Salivary Cortisol Enzyme Immunoassay Kit; Salimetrics LLC). A Tecan robot method was used, modified to suit a 25-μL saliva volume. The intra assay coefficient of variation (CV) was 5%, and the total CV was 7.7 and 7.5% for low and high values, respectively (32 (link)).
6
Salivary Cortisol and Testosterone Assessment
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Salivary samples to assess cortisol and testosterone levels were collected by passive drool. Participants were asked to deposit 5 ml of saliva in plastic vials, and samples were frozen at −80°C. Biochemical analyses were done by the Laboratory of Social Cognitive Neuroscience (Faculty of Psychology at the University of Valencia). For cortisol levels, the samples were analyzed in duplicate with the salivary cortisol enzyme‐immunoassay kit from Salimetrics. Assay sensitivity was 0.007 μg/dl. For each subject, all the samples were analyzed in the same trial. The mean inter‐ and intra‐assay coefficients of variations were all below 10%. For testosterone levels, the samples were analyzed in duplicate using enzyme‐immunoassays with the expanded range salivary testosterone enzyme‐immunoassay kit from Salimetrics. Assay sensitivity was 1.0 pg/ml, and the mean inter‐ and intra‐assay coefficients of variation were all below 10%.
7
Saliva and Nail Cortisol Quantification
8
Salivary Cortisol Measurement Using Salivettes
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The saliva samples were collected using salivettes (Sarstedt, Nümbrecht, Germany). To measure cortisol levels, salivary samples were analyzed in duplicate with the salivary cortisol enzymeimmunoassay kit from Salimetrics (Newmarket, UK). Assay sensitivity was 0.007 µg/dL, and the intra- and inter-assay variation coefficients were below 10%.
Participants were thoroughly instructed about how to provide saliva samples, and they were given written instructions. They were instructed to drink only water and not eat or brush their teeth at least 1 h prior to each saliva sample. To objectively verify participant adherence to the saliva sampling time at home, salivettes were stored in Medication Event Monitoring System (MEMS) TrackCap containers (MEMS 6 TrackCap Monitor, Aardex Ltd., Switzerland), and participants recorded the time of each saliva collection on a log.
Participants were thoroughly instructed about how to provide saliva samples, and they were given written instructions. They were instructed to drink only water and not eat or brush their teeth at least 1 h prior to each saliva sample. To objectively verify participant adherence to the saliva sampling time at home, salivettes were stored in Medication Event Monitoring System (MEMS) TrackCap containers (MEMS 6 TrackCap Monitor, Aardex Ltd., Switzerland), and participants recorded the time of each saliva collection on a log.
9
Salivary Cortisol Measurement in Infants and Mothers
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Salivary cortisol was obtained from infants and mothers 5 min before each challenge and 20 and 40 min post-challenge. Two Sorbettes (Salimetrics, State College PA) were collected for each participant at each time point. Saliva samples were centrifuged for 10-min at 3000 rpm to extract the saliva and then stored in a freezer at −70 °C. Salivettes were thawed and centrifuged for 10 min at 3000 rpm at 4 °C. All samples were assayed twice using a salivary cortisol enzyme immunoassay kit (Salimetrics, State College, PA), and average values were used in analyses. The interassay variability was 10.6%; the intraassay variation was 8.3%, for samples with low values, and 6.9% for samples with high values.
10
Salivary Cortisol Concentration Analysis
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After participants returned the saliva samples to the laboratory, the samples were kept in the refrigerator until they were centrifuged at 3,000 rpm for 5 min, resulting in a clear supernatant of low viscosity that was stored at −80°C until the analyses of the salivary cortisol levels. HPA axis activity was measured by analyzing the salivary cortisol levels. Salivary cortisol concentrations were determined in duplicate with the salivary cortisol enzyme immunoassay kit from Salimetrics (Newmarket, United Kingdom). Assay sensitivity was <0.007 μg/dl. For each subject, all the samples were analyzed in the same trial. The inter- and intra- assay variation coefficients were all below 10%. Cortisol levels were expressed in no/L.
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