Human igf 1 quantikine elisa kit

After overnight fasting, venous blood samples are taken from participants following gained consent for attaining blood sampling components. All blood samples are collected between 7:00 and 8:00 am to minimize the effects of diurnal variation of plasma BDNF levels [46 (link)]. Breakfast is prepared under the guidance of the nutritionists to ensure good compliance. Two nurses perform the blood sampling from cubital veins using EDTA-coated tubes. Blood samples are immediately placed on ice and then centrifuged at 3500 rpm for 15 min in a 4°C centrifuge. Plasma samples are collected and stored at −80°C before the measurement. The plasma IGF-1 and BDNF concentrations are measured in duplicate using Enzyme-linked Immunosorbent Assay (ELISA) kits (for the first assessment wave, Human IGF-1 Quantikine ELISA kit and Human BDNF Quantikine ELISA kit, R&D Systems, USA), according to the manufacturer's instructions.

CAF were cultured alone or in presence of cancer cells in serum-free medium. Supernatants were collected after 72 h and IGF-1 was measured by ELISA, using human IGF-1 Quantikine ELISA kit (R&D Systems) according to the manufacturer instructions. All IGF-1 values were normalized to amount of total proteins in the supernatant.

In MDD patients and HC, IGF-1 concentration was measured in the morning under fasting conditions. Blood was drawn by a venous puncture between eight and eleven a.m. within the first 2 days after clinical assessments in 125 participants at baseline and 48 patients after 2 months of vortioxetine (10–20 mg per day) treatment. Blood was centrifuged at 3000 g for 10 min, and the serum was stored at − 20 °C until further processing. IGF-1 was measured using the chemiluminescence immunoassay Immulite 2500 (Siemens AG, Germany) and Human IGF-1 Quantikine ELISA Kit (R&D Systems Inc., Minneapolis, USA). As IGF-1 standard, the WHO IRR 87/518 was used, which offers a measurement range with the Immulite test from 20 to 1600 ng/ml.

IGF-1 levels were measured using Human IGF-1 Quantikine ELISA Kit (R&D Systems; Cat. No.: RD-DG100). EDTA-plasma samples were collected from patients at the beginning and after the 12th week, the samples were stored at − 74 °C until performing the assay. The assay employs a quantitative sandwich immunoassay technique. The IGF-1 assay protocol was carried out according to the manufacturer’s instructions.

In MDD patients and HC, BDNF, and IGF-1 concentrations were evaluated in the morning. Blood was taken by a venous puncture between 8:00 and 11:00 a.m. within the first 2 days after clinical assessments in 73 participants at baseline and 30 patients after 8 weeks of vortioxetine (10‑20 mg per day) treatment. Blood was centrifuged at 3000 g for 10 min, and the serum was stored at -20° C until further processing. IGF-1 was assessed with the chemiluminescence immunoassay Immulite 2500 (Siemens AG, Germany) and Human IGF-1 Quantikine ELISA Kit (R&D Systems Inc., Minneapolis, United States). The measurement range for IGF-1 was from 20 to 1600 ng/ml. BDNF was measured using the chemiluminescence immunoassay «Sunrise» (TEKAN, Austria, GmbH) and BDNF Sandwich ELISA Kit («Millipore-ChemiKineTM»). The measurement range was from 15 to 1000 pg/ml.

We selected the ligands, such as EGF, TGF-alpha, AREG, EREG, NRG, HGF and IGF-1, that had previously been reported to be associated with activation or cross-talk of the downstream signal pathway of the ErbB family in solid tumors. We used ELISA kits to measure serum levels of ligands as follows: Human HGF Quantikine ELISA Kit (DHG00, R & D Systems, Minneapolis, MN, USA), Human epiregulin ELISA kit (CSB-EL007779HU, CUSABIO, Wuhan, Hubei, China), Human amphiregulin ELISA kit (E90006Hu, Uscn Life Science, Wuhan, Hubei, China), Human EGF Quantikine ELISA kit (DEG00, R & D System, Minneapolis, MN, USA), Human TGF-alpha Quantikine ELISA kit (DTGA00, R & D System, Minneapolis, MN, USA), Human Neureglin-1 ELISA kit (CSB-E17153 h, CUSABIO, Wuhan, Hubei, China) and Human IGF-1 Quantikine ELISA kit (DG00, R & D System, Minneapolis, MN, USA). Protocols for ELISA of these ligands are summarized in Supplementary Material 2.