No Bands on Western Blot

[clokke76]

Hi,I am new here.

I am getting no bands on my western

what should i do?

[admin]

Ok will look into this.

[danfive]

1. Check transfer efficiency, stain used gel after transfer, or protein blot on membrane.
2. Check any reference material to see if you should be running denatured or native proteins.
3. If you are running protein mix, like a cell lysate, what percent or amount should be your target protein.
4. Troubleshoot the Antibody concentration by running the positive control in multiple lanes, blotting, cutting membrane into strips and blotting one condition on strip at a time until find best conditions.
5. Check your detection reagents.
6. Check your camera or scanner. (increase exposure time)
7. You can even try blotting by skipping the blocking step, but expect to see nonspecific binding at areas of high protein concentration though. You may see a very weak signal this way at your expected size range though.