ELISA was performed using human VEGF Quantikine PharmPak kit for VEGF‐A (R and D systems, USA), by following the manufacturer's instructions. 100 μL of CMs was added to 96‐well microwells and incubated for 2 hours at room temperature. After washing with PBS, 100 μL of biotin‐conjugate was added and washed. Next, 100 μL of streptavidin‐HRP was added to each well and kept for 1 hour at room temperature, and 100 μL later mixed and incubated for 30 minutes. The reaction was terminated by the addition of 100 μl of terminating solution. Colour density was read at 450 nm using a microwell reader (BioTek, Winooski, VT, USA).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.