So these are my saline-treated cerebellum samples. Lanes 1-9 are loaded with equal amount of protein. This membrane was stained for b-actin only, and as you see some bands are missing (my protein of interest, which is 96kDa is absolutely fine). What happened to some of my samples? Transfer looks fine

Could it be that lower MW protein just went missing during homogenisation/heat treatment?

For reference, all samples were processed on the same day using the same buffers. Also, my treated samples are absolutely fine.

10ug protein per well

5% milk

beta actin 1:5000

seconday Ab 1:2000

Thanks