Last updated: 1/12/12 By: Tim Starr
Instead of running protein sample on gel and transfering to membrane, you can just place a drop of protein directly onto the membrane. Do this to test whether your antibodies can detect the protein. The following protocol is taken from the Qiagen Bench Guide.
- Dilute protein samples in dilution buffer to final protein concentration of 1 to 100 ng/µl.
- Place 1 µl on a dry membrane (Qiagen recommends nitrocellulose).
- Let the sample dry for a short time at room temperature.
- Alternatively, can apply crude cell lysate (1 µl) if you think the concentration is 1 to 100µl ( I assume this is the concentration of the protein you wish to detect, not total protein concentration).
- Follow protocol for detecting proteins using antibodies
Dilution buffer 1 x, 1 L (denaturing conditions) adjust pH to 8.0