Bio-Rad. Troubleshooting Western Blots with the Western Blot Doctor™

The Western Blot Doctor is a self-help guide developed by Bio-Rad researchers that enables you to identify and troubleshoot western blotting problems. Comprehensive solutions and suggestions are provided to help solve your particular western blotting challenges.with the Western Blot Doctor™ 

Tips, Techniques, and Technologies from the Western Blotting Experts at Bio‑Rad Laboratories:  Get Your Free Guide

The Western Blot Doctor is a self-help guide developed by Bio-Rad researchers that enables you to identify and troubleshoot western blotting problems. Comprehensive solutions and suggestions are provided to help solve your particular western blotting challenges.

To start troubleshooting your western blotting problems, choose the type of problem you are experiencing.

I am having trouble with:

Protein Band Appearance

• Inconsistent control protein levels across samples
• Ghost protein bands
• Swirls or missing bands; bands appear diffuse on blot
• Blurry bands

• Bands are curved (smiling) not straight
• Broad or misshapen bands
• White (negative) bands on film using ECL method
• No bands

Signal Strength

• Uneven signal across blot
• Saturated band signal
• Faint high-MW bands
• Faint bands, weak or no signal

Protein Band Size and Pattern

• Band(s) at lower MW than expected
• Band(s) at slightly higher MW than expected, and may be blurry
• Band(s) at significantly higher MW than expected
• Multiple bands at various MWs

Blot Background

• White spots or regions on membrane
• High background on blot
• Blotchy or patchy background
• Uneven spots on blot or speckled background
• Shadow of tank blotter gel cassette on blot

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