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Similarly, Rajegowda et al

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

The proteome concentration of the tissue lysates was determined using the DC protein assay (Bio-Rad), normalized to 2 mg/mL, and then 100 L of each sample was transferred to a LoBind Eppendorf tube containing 48 mg of urea