The melatonin content in cell supernatants was measured using the competitive inhibition ELISA kit (CEA908Ge, USCN Life Science INC, China) according to the manufacturer’s protocol. First, 50 μL of each sample dilution was added into the wells and incubated with 50 μL of detection reagent A for 1 h. Then, 100 μL of detection reagent B was added and incubated for 30 min at 37°C. Next, 90 μL of the substrate solution was added and incubated for 15–25 min at 37°C. The reaction was stopped with 50 μL of stop solution, and optical density (OD) values were immediately measured using an ELISA analyser at 450 nm (Bio-Rad, Model 680).
Cea908ge
Manufactured by USCN
Sourced in China
The CEA908Ge is a laboratory instrument designed for the detection and analysis of various analytes. It utilizes electrochemical techniques to facilitate measurements. The core function of this product is to provide accurate and reliable data for research and diagnostic applications.
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Lab products found in correlation
2 protocols using cea908ge
1
Melatonin Quantification in Cell Supernatants
2
Melatonin Signaling in Avian Cells
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All of the blood samples were heparinized with 1000 UI/mL of sodium heparin (H4784, Sigma, St. Louis, MO, USA) in avian saline. After centrifugation at 1000× g for 20 min, the plasma was decanted and stored at −80 °C. Concentrations of MEL in plasma were measured by an enzyme-linked immunosorbent assay kit for anti-MEL (CEA908Ge, Uscn Life Science, Inc., Wuhan, China).
The cells were incubated with 1 μM of prazosin for 30 min followed by 250 pg/mL of MEL for 24 h, and 3-isobutyl-1-methylxanthin (IBMX 100 μM, T1713, Topscience, TX, USA) was added to the medium to prevent cAMP degradation before the cells were incubated for a further 30 min. The cells were collected and lysed with lysis buffer from the cAMP ELISA Kit (R&D Systems, Minneapolis, MN, USA). Further treatment of the samples was performed according to the manufacturer’s instructions.
The cells were incubated with 1 μM of prazosin or 10 μM of PD98059 for 30 min, followed by 250 pg/mL of MEL for 24 h. The supernatant was collected for IGF-I protein detection by the anti-chicken IGF-I ELISA kit (SEA050Ga, Uscn Life Science, Inc., Wuhan, China), according to the manufacturer’s protocols.
The cells were incubated with 1 μM of prazosin for 30 min followed by 250 pg/mL of MEL for 24 h, and 3-isobutyl-1-methylxanthin (IBMX 100 μM, T1713, Topscience, TX, USA) was added to the medium to prevent cAMP degradation before the cells were incubated for a further 30 min. The cells were collected and lysed with lysis buffer from the cAMP ELISA Kit (R&D Systems, Minneapolis, MN, USA). Further treatment of the samples was performed according to the manufacturer’s instructions.
The cells were incubated with 1 μM of prazosin or 10 μM of PD98059 for 30 min, followed by 250 pg/mL of MEL for 24 h. The supernatant was collected for IGF-I protein detection by the anti-chicken IGF-I ELISA kit (SEA050Ga, Uscn Life Science, Inc., Wuhan, China), according to the manufacturer’s protocols.
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