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
Soluble FN1 protein levels secreted by HEK293 cells were determined using a human FN1 enzyme-linked immunosorbent assay (ELISA) (Boster Biological Technology). The assay was performed according to the manufacturer’s instructions. HEK293 cells were transfected with the indicated siRNAs for 72 h and starved with fetal bovine serum free Dulbecco’s modified Eagle’s medium for 16 h before the collection of the cell culture supernatant. Fibronectin protein amounts were normalized to cell numbers determined by flow cytometry.
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