I'm currently facing a challenge with c-fos staining, and I've made several attempts to resolve the issue. However, even after trying different antibodies, I consistently observe the same staining patterns. Specifically, on the right side, the staining appears to be concentrated within the nucleus, while on the left side, the staining seems to be displaced from the nucleus. I'm uncertain which staining pattern represents the genuine signal and which one might be an artifact ?
Hello Yaseen, I think twice wether your Antiboy is specific. Your imgages show alot of unspezific background staining. I recommad 2 things. 1. use suitable positiv controls so that you know your protocol and your antibodies are working properly. 2. Use instead of a green fluorescence dye like Cy2 or Alexa 488 a red one like Cy3 or Cy5. In the attachment you will find 2 images of the cfos antibody from Synaptic Systems . 1. IF image shows cfos in red (Cy5 labled secondary antibody). 2. brightfield image cfos detected with DAB. Both images (brain cortex) show a clear nuclear staining of the cfos expressing cells.
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