Hi all,
I have a question about my lip3000 transfect neurons. I transfected my primary neurons with a control(mRFP) plasmid. Then I did immunostaining with primary proteinX antibody and Alexa 488 secondary antibody. Interestingly, for my control group, the neurons that were transfected with mRFP plasmid showed a very low signal of green fluorescence; for the neurons that were not transfected with mRFP, the GFP signal is normal. I am curious why it will happen? Ideally, it shouldn't see the difference between transfected cells and nontransfected cells. I just feel like the cells that are transfected are not likely to be immunostained. Did anyone have the same issue as mine? Will the lip3000 affect the permeability? I used 0.2% TriX-100 for 5min.
Thank you so much!
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