proACE™ Ultra ECL direct

Protocols

Prior to Use:

Prepare your protein blot as usual. After incubation with HRP-conjugated secondary antibody, wash 3x 5min and leave membrane in wash buffer in order to prevent drying until ready to proceed with development.

For step 2, approximately 0.1ml of proACE™ Ultra ECL direct is required per cm2 of membrane. Typically, for a mini-sized membrane, 4 ml is sufficient. 

Usage 

1. Transfer the membrane, with the protein side up, to a clear and level surface or into a clean container
2. Cover the membrane with proACE™ Ultra ECL direct and incubate for 90 seconds at room temperature (The amount of time required may be optimized individually in order to obtain stronger signal as it depends on the amount of secondary antibody bound. In general, 30 seconds to 2 minutes is more than enough as proACE™ Ultra ECL direct is extremely sensitive).
3. Remove the membrane from the mixture, drain of any excess and cover the damp blot with plastic wrap to prevent from drying.
4. Image by exposing to X-ray film or CCD imager. For an unknown signal, try to expose for 15 seconds, 30 seconds, 1 minute and 5 minutes for initial evaluation.