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Download troubleshooting handbooks for IHC, Western blot and ELISA for FREE.
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Western Blot Tips & Tricks: Blocking Optimization | ||||||||||||||||
Greetings Earthling, Following protein transfer, it is important to block the unreacted sites on the membrane using inert proteins and/or nonionic detergent to reduce levels of nonspecific protein binding during the assay. Blocking buffers should block all unreacted sites without disrupting target protein-membrane interactions or affect epitope availability. There are three factors to consider when choosing an appropriate blocking agent for your specific protocol: (Click the meme for blocking optimization tips!)
The most typical blocking solutions are nonfat dry milk, casein, gelatin, or Tween-20 in TBS and/or PBS buffers. Here are some advantages and disadvantages of some common blocking agents:
We advise reviewing the data sheet for any recommended blocking agents because antibodies can be very sensitive to the blocking agents used. Here is a summary workflow for a typical WB experimental protocol along with necessary reagents for each step. Click on the flowchart image to shop WB Reagents & Kits, 20.17% off for a limited time! Many different factors in the blocking step may need troubleshooting during western blotting... For more troubleshooting and optimization tips, visit our technical resource center: |
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More WB Resources | ||||||||||||||||
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Featured Western Blotting Products | ||||||||||||||||
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WB Blocking Reagents | ||||||||||||||||
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