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  • Labeling CLIP-tag Purified Protein In Vitro (P9311)

    Protocol

    1. Thaw the CLIP-tag Purified Protein or CLIP-tag fusion protein at room temperature just before use.

    2. Set up the reactions, in order, as follows:
      Component Volume Final
      Concentration
      Deionized Water 32 μl  
      5X CLIP-tag
      Reaction Buffer
      10 μl 1X
      50 mM DTT 1 μl 1 mM
      50 µM CLIP-tag
      Purified Protein
      5 μl 5 µM
      250 µM CLIP-tag
      Substrate
      2 μl 10 µM
      Total Volume 50 μl  
    3. Incubate in the dark for 60 minutes at 37°C. 

    4. Run sample on an SDS-PAGE gel and detect using a fluorescent gel scanner or store samples at -20°C or -80°C in the dark.


      Notes for In Vitro Labeling

      Adjust the volumes to achieve the final concentrations indicated if the stock solutions of protein, substrate, and DTT are different than described above. 

      One may choose to label the CLIP-tag Purified Protein or CLIP-tag fusion protein in a different final concentration, however, it is recommended to use a 2-fold excess of CLIP-tag substrate vs. CLIP-tag protein. 

      Labeling may also be carried out in 1X PBS solution or 50 mM HEPES containing 1 mM DTT.