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Nano-Glo HiBiT blotting

DR Deshani C. Ranawakage
TT Takuya Takada
YK Yusuke Kamachi
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To detect the HiBiT-derived signal from the GST proteins fused with monomeric, dimeric and trimeric forms of the epitope tags and the HiBiT peptide, equal amounts of the purified proteins were separated on 10% SDS-polyacrylamide gels and transferred to nitrocellulose membranes. The protein-transferred membranes were incubated in TBST for 30 min, and this medium was then replaced with Nano-Glo HiBiT blotting reagent containing LgBiT protein (Promega). After 1 hour of incubation at RT, the substrate furimazine was added, and the incubation was continued for another 5 min. The blot was imaged using a chemiluminescence imager with a CCD camera (Fusion, Vilber Lourmat).

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