I've tried using PBS and carbonate buffer to coat. Tried 2 hours at RT as well as overnight at 4C. GFP washes away after washing with PBS/.05%Tween 20. Any ideas how to coat GFP to the plate?
Hi Kristina,
Try using high binding ELISA plates.
And washing with tween buffer is not recommended at the coating step.
You may simply wash with PBS/carbonate buffer.
Here is a technical guide for more information.
https://www.seracare.com/globalassets/seracare-resources/tg-protocols-and-troubleshooting.pdf
Best,
S
Sudeep is right, high binding plates in combination with carbonate buffer should do the trick. Good luck!
Use Immubilon high protein Bing protein and while performing the first step of coating GFP prepared in carbonate-bicarbonate keep the buffer pH at 9.3 to 9.6 to impart a net negative charge on GFP. Keep GFP coated ELISA plate overnight at 8C for optimal binding of GFP. If PBS doesn't work try using coating buffer for washing, too.
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