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HiBiT Blotting: A Fast and Sensitive Western Blot Alternative

  • Results in as little as 5 minutes
  • No antibody required
  • Low background and high sensitivity
  • No washing or blocking steps

Compare Protocols

See how HiBiT blotting compares to traditional FLAG methods

Request Application Note

What Is HiBiT Blotting?

HiBiT is a bioluminescent detection method based on protein complementation.

The HiBiT tag is added to your protein of interest and detected when the complementary LgBiT subunit binds to HiBiT, forming a functional luciferase enzyme.

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HiBiT Is Different from Traditional Epitope Tags

  • In traditional Western blotting, you need an antibody to your protein of interest or epitope tag, and a method for detecting that antibody.
  • With HiBiT, no antibody is needed, eliminating processing steps and minimizing hands-on time.

Benefits of HiBiT Protein Tagging

  • Easy quantitation and detection
  • No blocking needed
  • No background, unlike antibody-based methods, with HiBiT blotting it doesn't matter if the detection reagent binds non-specifically to the membrane. There will be no luminescent signal except where it has bound to a HiBiT tagged protein.

How HiBiT Tagging Is Being Applied

  • Monitoring protein abundance in the cell
  • Protein degradation research
  • Monitoring receptor internalization
  • Inserting and detecting CRISPR knock-ins

See the Data

Download the application note, or visit the Nano-Glo® HiBiT Blotting System product page to learn about the sensitivity and time-saving advantages of the HiBiT tag compared to traditional blotting methods that use an antibody to an epitope tag (e.g., FLAG tag).