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However, some data was missing among the protein concentrations

This is what it looked like for me My work is research, writing, and teaching

Assuming you followed the above advice and used an appropriate binding buffer and your protein isnt huge, then a likely reason for this is that the expression yield is much lower than you expected

Reference ranges vary by lab, but most experts agree that: Total testosterone: roughly 3001,000 ng/dL is considered normal Free testosterone: about 50210 pg/mL is considered normal Anything consistently below 300 ng/dL for total testosterone is often classified as hypogonadism, or clinically low testosterone