To reduce background noise on your Western Blots, try increasing the number of washes (e.g., 5 washes for 5 mins each) and adding 0.1% Tween-20 to your TBST wash buffer. A fresh blocking solution also helps! Using high-quality antibodies is key.

For tricky cell lines that lift off during staining, coat your glass coverslips with Poly-L-Lysine (0.1 mg/mL in water) for 1 hour at room temperature before seeding. This provides a positively charged surface that promotes adhesion. Remember to wash with sterile water before adding cells.