Western Blot Protocol Beginner Tutorial with Operations

severe

Band smearing / streaking

Bands have blurred edges, show vertical smearing or the entire lane is hazy; background is fogged, making band intensity comparisons unreliable.

💡 4 · ✓ 4

moderate

Low Acrylamide Percentage for Target Protein Size

Lower molecular weight proteins migrate excessively fast through the gel, resulting in poor resolution and bands appearing at the bottom of the membrane or running off entirely.

💡 3 · ✓ 4

moderate

Black Dots or Speckled Background on Western Blot

Non-specific dark dots or speckled pattern appear across the membrane background, not localized to protein bands. The signal appears as discrete spots rather than uniform background.

💡 5 · ✓ 5

minor

Smiling bands (curved bands)

Bands curve upward at the edges of the gel, appearing like a smile — the middle of the lane runs faster than the sides.

💡 4 · ✓ 4