I am looking to use ROSA mTmG mice with a cell-specific Cre. I will have tissues fixed in 4% PFA, paraffin-embedded, and sectioned for studies. Is the signal from tdTomato and GFP strong enough that I don't need to come in with an antibody for immunofluorescence staining?
I am interested in staining for other protein targets on these tissues. I'm wondering whether the signal is too strong in the green and red channels from this mouse and if this will make staining other targets difficult, as I wouldn't be able to use the red/green channels as readily. Thanks!
Hi interesting question. I recently looked at the cre tissue specificity, using the tdTomato reporter system. I believe there are a number of potentials issues that depend on tissue processing. I was informed that fixation in 4%PFA and paraffin embedding substantially reduced the fluorescent signal, I was advised to use the antibody detection system to detect positivity if I used this system. However I embedded the tissue in OCT for frozen tissue sectioning, this preserved the fluorescent signal. Very strong signal. I would check the intensity of the signal in the cut sections before proceeding with any IHC.
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