Flag-tagged protein on western blot ?

16 February 2017 1 5K Report

Hi,

Can anyone tell me what to expect to see on a western blot of a FLAG-tagged protein? the one i'm working on is FLAG-VDAC1 so do I see one 1 or 2 bands? i'm using a GST primary antibody.

Many thanks!

Laura Torrente

I think you are a bit confused...

Flag is a tag (small peptide) that you add to your protein (either to the N or C-terminus) to facilitate the detection or pull down of your protein. The number of bands that you are going to see does not depend on your tag, depends on the protein you are studying. Flag is a very small peptide, so I would not expect variations on the original molecular weight of your protein of interest (VDAC1)

I do not understand why would you use a GST antibody to detect it... unless your protein is Flag and GST-tagged...

I would recommend you to do some reading before you do these assays, specially check papers that they have been working before with this protein.

Good luck!

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