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. 2023 Jan 4;4(1):101992. doi: 10.1016/j.xpro.2022.101992

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© 2022 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Testing biotin labeling of PKAc-miniTurbo clonal line

Western blot of clonal cell line lysates in 4 conditions probing with NeutrAvidin™-HRP at 1:4000 dilution. Protein biotinylation by PKAc-miniTurbo occurs only when bait protein expression is induced (+ doxycycline) and exposed to exogenous biotin (+ biotin) as shown in lane 4. Bands at ∼65 kDa and ∼130 kDa in lanes 1–3 are endogenous biotinylated proteins. Ponceau S demonstrates equal protein loading. Doxycycline treatment was 48 h. Biotin incubation was 2 h.