I'm doing western blot, using some drug,

All condition and process are same.

But only "V" drug treatment groups are show different result in western blot band include beta actin.

1. While other groups show normal results, "V" drug treatment group (3th, 4th band in picture) was not visible or weak(File.actin2) or spread(File.actin1,2).

2. I confirmed TAZ antibody in PVDF membrane, antibody position was not visible or different position(File.TAZ)

I use Ripa buffer and proteinase in cell prep, and set total protein concentration using Bradford assay,

Running : 10% Tris-glycine gel, 1mg/ml protein loading, 100~110V for 90min

Transfer : PVDF membrane, 90V for 60~90min in ice

Was something interfere when protein prep or run in gel ?

Anyone Who has similar experience, please help..