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
See how HiBiT blotting compares to traditional FLAG method in this paper
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.
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).
Read more about HiBiT blotting applications in these references:
- Nakashoji, A. et al. (2020) Identification of a Modified HOXB9 mRNA in Breast Cancer. Journal of Oncology. Article ID 6065736,
- Schwinn, M.K. et al. (2020) A Simple and Scalable Strategy for Analysis of Endogenous Protein Dynamics. Sci. Rep. 10(1), 8953.
- Tange, N. et al. (2020) Staurosporine and venetoclax induce the caspase-dependent proteolysis of MEF2D-fusion proteins and apoptosis in MEF2D-fusion (+) ALL cells. Biomed Pharmacother. 128, 110330.
- Ranawakage D.C. et al. (2019) HiBiT-qIP, HiBiT-based quantitative immunoprecipitation, facilitates the determination of antibody affinity under immunoprecipitation conditions. Sci. Rep. 9(1) 6895.
- Sasak, M. et al. (2018) Development of a rapid and quantitative method for the analysis of viral entry and release using a NanoLuc luciferase complementation assay. Virus Res. 243, 69-74.
- Tamura, T. et al. (2019) In Vivo Dynamics of Reporter Flaviviridae Viruses. J Virol. 93(22), e01191-19.