During western blotting, the protein ladder transferred to the membrane but not my sample protein, why not?

Question

During western blotting, the protein ladder transferred to the membrane but not my sample protein, why not?

Answer 1

You need to start with some trouble shooting. Did you measure the protein concentration of your sample before you loaded it? Sometimes the protein gets degraded before you run the sample. Also I would you ponceau S stain (its a red stain) to check and see if any faint proteins bands appear in your sample lane.

If those are normal, then you may have an unusual protein that transfers poorly. If your protein is very large then you may have to transfer longer, if your protein is smaller then you may have to transfer a short amount of time.

Good Luck.