what are wildtype control, non-silencing control and control? - (Jun/23/2015 )

Hi guys,

i have a simple question but confused me. Let's say i am doing western blot.

If i have an immortalized cells (using Htert) overexpressing Akt, what should be the control cell for comparison?

Q 1. Can it be like this?

control - immortalized cells expressing GFP construct

OE cells- immortalized cells overexressing protein of interest (Akt)

Q 2. or can the control be just immortalized cells as they already have Htert construct?

For downregulation, i know the control is generally a non-silencing construct.

But what do you use as a control against overexpressing cell ?

thanks a lottt

In my humble opinion, your control can be simple immortalized cells. Anything that won't show a big fat band for Akt in a WB actually. So to answer Q1: yes and Q2: yes again.

Good luck !