I m using immunoflourecense stain for detection of MyoD and Myogenin expression in murine skeletal muscle injury model, and I got cytoplasmic signal in most of regenerating myoblasts, should I consider this signal as a false positive because what I know that Myod and Myogenin is highly expressed in nuclei of myocytes. Note, I got some signal in nuclei too.
I use donkey serum 10% to block my sections and I do use non-injured muscle as negative control.
Hi!
I use to block with 5%BSA (igG free) + 10% goat serum in PBS 1X solution . Moreover, if the primary antibody is an anti-mouse I perform a second blocking step with Fab blocking solution (M.O.M).
I am also trying to immunolocalize myogenin in regenerating skeletal muscle (7dpi). I expected to found many central nuclei positives for MyoG but I have almost all positive label in peripheral nuclei...Does that make sense to you? Thank you in advance.
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