Western Blot background GFP bands-- how did you reduce this in WTC-11?

hiPSC Culture Discussion
1

Hi there,

From your Roberts et al., 2017 pub, your GFP Western Blots look very clean, but I’m using the same antibody specified and getting so many background bands in both WT and my mEGFP-tagged line lysates. Any chance someone could help me consider what might be going on or offer some tips with this particular antibody/cell type combo for my Westerns?

Thank you very much!

Maggie Fuqua

2

Hey Maggie! Apologize for the delay, but I got an answer from our team I’m pasting below. Let us know if it helps!

“Assuming that you are collecting lysate from hiPS cells, I suspect your background may be due to Matrigel in the system. To minimize this, we dissociate cells from the Matrigel substrate before lysis and sample prep for western blots.”