WB Technical Resources

Protocols, optimization tips,
troubleshooting guides,
and more for Western Blot.

Troubleshooting guides

Troubleshooting guides

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handbooks for IHC, Western
blot and ELISA for FREE.

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Western Blot Troubleshooting Weak Signal

Weak signal is typically caused by problems in the blocking or washing steps, but can also be caused by a number of other issues. Use these tips to identify and troubleshoot you weak signal results:

Keywords: Weak Signal, Low Signal, Western Blot Troubleshooting

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Western Blot Troubleshooting Bands Wrong Molecular Weight

Western blotting separates proteins based on size; large proteins migrate through a polyacrylamide gel matrix slower than small proteins do. However, other factors can influence the migration rate of proteins, resulting in band sizes different than predicted based on protein size alone.

Keywords: Band Size, Wrong Band Size, Western Blot Troubleshooting

Learn More About Band Size

Western Blot Troubleshooting Background Background Blotchy, Flecked, Or Dirty

Many Western blot problems arise due to issues with the background. Splotches, streaks, or background staining can ruin results. Use these troubleshooting tips to identify and resolve the cause of your background troubles:

Keywords: Flecked Background, Speckled Background, Blotched Background, Western Blot Troubleshooting

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Western Blot Troubleshooting High Background

High background on a western blot occurs when the background signal of the membrane reduces the signal-to-noise ratio to unreadable levels. Use these troubleshooting tips to identify and resolve the cause of your high background:

Keywords: High Background, Saturated Background, Background Staining, Nonspecific Staining, Western Blot Troubleshooting

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Western Blot Troubleshooting Distorted Bands

Distorted bands can make it very hard to interpret your results. Common distortions include smile shaped bands with the edges trailing upward, diffuse bands that are broad or blurry, and streaked bands that trail off in several directions. Make sure your next blot has even, crisp bands by following these tips:

Keywords: Smile Bands, Distorted Bands, Uneven Bands, Western Blot Troubleshooting

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