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tyramide azide  (Iris Biotech Gmbh)
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Msm expressing APEX2, Sec-APEX2 or Tat-APEX2 was grown without or with theophylline and subjected to the labeling protocol with either <t>tyramide</t> <t>azide</t> or tyramide alkyne. Lysates underwent a CuAAC reaction with alkyne or azide-conjugated fluorescein. In-gel fluorescence detection of the clicked lysates was used to assess protein labeling. Equal protein loading was confirmed by subsequent Coomassie staining (Figure S5). Data are representative of 3 biological replicates under the specified conditions.
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Msm expressing APEX2, Sec-APEX2 or Tat-APEX2 was grown without or with theophylline and subjected to the labeling protocol with either tyramide azide or tyramide alkyne. Lysates underwent a CuAAC reaction with alkyne or azide-conjugated fluorescein. In-gel fluorescence detection of the clicked lysates was used to assess protein labeling. Equal protein loading was confirmed by subsequent Coomassie staining (Figure S5). Data are representative of 3 biological replicates under the specified conditions.

Journal: ACS infectious diseases

Article Title: Compartment-Specific Labeling of Bacterial Periplasmic Proteins by Peroxidase-Mediated Biotinylation

doi: 10.1021/acsinfecdis.8b00044

Figure Lengend Snippet: Msm expressing APEX2, Sec-APEX2 or Tat-APEX2 was grown without or with theophylline and subjected to the labeling protocol with either tyramide azide or tyramide alkyne. Lysates underwent a CuAAC reaction with alkyne or azide-conjugated fluorescein. In-gel fluorescence detection of the clicked lysates was used to assess protein labeling. Equal protein loading was confirmed by subsequent Coomassie staining (Figure S5). Data are representative of 3 biological replicates under the specified conditions.

Article Snippet: Biotin-phenol (Iris Biotech), tyramide azide and tyramide alkyne were prepared as 50 mM stocks in DMSO and used in cultures at 1 mM final concentration (final DMSO concentration was 2%).

Techniques: Expressing, Labeling, Fluorescence, Staining