I am looking for transporter protein expression on cancer cell lines. Do I need to use transporter protein over-expressing cell line as positive control for my western blot experiment? for example if I am looking at OCT expression on particular cancer cell line means do I need to also use some OCT over expressing cell line as positive control? is it mandatory? and why? Already I am getting decent results with OCT protein expression on cancer cell lines...
Hi there,
This control is necessary to show clearly where is the band of interest on the membrane (by comparing this positive control to the negative control (ie. cells not expressing the protein)). It is also confirming that the WB is working OK or not.
Simply speaking control for western blotting is to check whether the system is working or not? If you are trying to detect your transporter protein with His-tagged, you can use any his tagged protein (need not tone a transporter protein, could be any soluble protein) in one of your lanes to check you get signal as a positive control. For negative control you always use your cell lines without your transporter protein expressed. By this way, you have a positive control and a negative control. Best wishes. Thamarai
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