I've been optimizing a Western protocol for phosphorylated proteins and have had issues with my red channel. Proteins in my green channel would not show up in my red channel. I would also have to turn down the red channel's intensity a lot more than the other channel because of how much background would be stained in red.

Has anyone experienced this? Any thoughts on how I might solve this? I've attached one of my blots in the red channel.

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