Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
-Store at 4°C for 6 months, at -20°C for 12 months.
-Avoid multiple freeze-thaw cycles.
-The standard solutions are best used within 2 hours. |
-The diluted Avidin-Biotin-Peroxidase Complex (ABC) solution should not be prepared more than 1 hour prior to the experiment.
-Prepare the substrate solution immediately before use or bring the pre-made substrate to room temperature.
-Sodium azide is an inhibitor of HRP. Do not include the azide in buffers or wash solutions if HRP-labeled conjugate is used for detection. |
|
Upstream tips |
-Store at 4°C for 6 months, at -20°C for 12 months.
-Avoid multiple freeze-thaw cycles.
-The standard solutions are best used within 2 hours. |
Protocol tips |
-The diluted Avidin-Biotin-Peroxidase Complex (ABC) solution should not be prepared more than 1 hour prior to the experiment.
-Prepare the substrate solution immediately before use or bring the pre-made substrate to room temperature.
-Sodium azide is an inhibitor of HRP. Do not include the azide in buffers or wash solutions if HRP-labeled conjugate is used for detection. |
Publication protocol
Culture media of 1×106 cells were collected separately after incubation for 72 h and then centrifuged at 10,000 × g for 10 min at 4°C. The supernatant OPN and IL-6 concentrations and the plasma OPN and IL-6 levels in patients with HNC were assessed using the Human OPN ELISA kit (Boster, China) and human IL-6 ELISA kit (Boster, China). Moreover, the mouse OPN ELISA kit (Boster, China) was used to measure the plasma OPN level of nude mice. The OPN and IL-6 concentration in culture media or plasma were calculated according to the formula of the standard curve. See Supplementary Materials and methods for detailed methods.
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Manufacturer protocol
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