How do I optimize the detection results with low molecular weight proteins?

To optimize low molecular weight protein , it is recommended to use a high-percentage gel to achieve optimal protein resolution. The percentage of acrylamide in the gel determines the pore size, which directly impacts protein separation based on size. High-percentage gels have smaller pores (ideally 0.2 μm) to ensure efficient transfer of small proteins (<20 kDa), making them suitable for separating smaller proteins more effectively. Another way to optimize the results is to use tricine-based acrylamide gels, since they offer higher resolution for low molecular weight proteins compared to glycine gels. Tricine's properties lead to more uniform stacking of proteins, preventing overloading and improving band visualization. One may also adjust transfer conditions such as voltage, transfer length, temperature, and buffer composition for efficient transfer of these proteins. Lower voltage and shorter transfer times are often used to prevent over-transfer.