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According to a report on GEN, 41% of researchers admit that their Western blots are unsuccessful at least 25% of the time.
Yikes! Western blotting (WB) is a widely practiced analytical technique to detect target proteins within samples using antigen-specific antibodies. When it fails to perform as expected, it can really be a downer.
We’re here to help you succeed. Next time you encounter another problem with Western blot, we’ve compiled a checklist to help you troubleshoot your experiment.
Cause | Solution |
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Antibody incubation temperature was too high |
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Antibody cross-reacted with other proteins or the blocking agent |
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Insufficient blocking |
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Insufficient washing |
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Cause | Solution |
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Insufficient sample loaded on the gel |
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Loss of primary antibody effectiveness |
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Inhibition of secondary antibody by sodium azide |
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Antigen masking by blocking buffer |
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Cause | Solution |
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Primary antibody concentration was too high |
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Excess protein on gel |
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Insufficient washing |
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Blocking problem |
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Access our Technical Resource Center for more WB tips and troubleshooting guidance.
*Note to educators: You are encouraged to share Boster Bio's resources and PDFs on your class and lab websites, please cite or link to origin bosterbio.com
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All Boster antibodies and ELISA kits are guaranteed to meet the specifications on the data sheet. We promise to thoroughly investigate any concerns about the quality of our products; if you encounter a problem with a Boster antibody or ELISA kit, our technical support team will respond with personalized advice within 24 hours. If we can’t make your experiment work, we will refund your purchase in full or provide a replacement free of charge.
Review : "This is an excellent antibody to endoplasmin in HC11 cells. Clean, reliable detection with very little if any background. Great antibody for Westerns. Could probably be optimized for 1 hour primary incubation."
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