Product FAQ

I want a metabolic label to be incorporated into proteins or other cellular structures by adding the reagent to the media of cultured cells or in vivo injection. What are my options?

Answer

There are three types of metabolic labeling reagents available:


  • Click chemistry (azide-alkyne)
  • Staudinger ligation (azide-phosphine)
  • Photoreactive amino acids

  • The incorporation of azide-modified biomolecules permits eventual detection or crosslinking using alkyne or phosphine reagents. The incorporation of alkyne-modified biomolecules allows eventual detection or crosslinking using azide-modified reagents. The incorporation of either EdU or EU requires that the cells/tissues possess a pyrimidine salvage pathway. 

    Answer Id: E15684

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    Product FAQ

    Can I perform Click-iT EdU detection on cells growing in 3D culture?

    Answer

    We have not validated the use of EdU for proliferation in 3D culture systems, but as this reagent is compatible for labeling cells in vivo, it is also expected to label cells in 3D culture systems. There are a number of reports in the literature that use this product in 3D culture systems; here are some citations:

    Lei Y, Schaffer DV (2013) A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation. Proc Natl Acad Sci U S A 110:E5039-E5048.
    Derda R, Laromaine A, Mammoto A et al. (2009) Paper-supported 3D cell culture for tissue-based bioassays. Proc Natl Acad Sci U S A 106:18457-18462.
    Robertson FM, Ogasawara MA, Ye Z et al. (2010) Imaging and Analysis of 3D Tumor Spheroids Enriched for a Cancer Stem Cell Phenotype. J Biomol Screen 15:820-829.

    Answer Id: E14987

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    Product FAQ

    Can I perform Click-iT EdU TUNEL detection on cells growing in 3D culture?

    Answer

    We have not validated the use of EdU TUNEL for apoptosis detection in 3D culture systems, but as this reagent is compatible for labeling cells in vivo, it is also expected to label cells in 3D culture systems. There are a number of reports in the literature that use this product in 3D culture systems; here are some citations:

    Lei Y, Schaffer DV (2013) A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation. Proc Natl Acad Sci U S A 110:E5039-E5048.
    Derda R, Laromaine A, Mammoto A et al. (2009) Paper-supported 3D cell culture for tissue-based bioassays. Proc Natl Acad Sci U S A 106:18457-18462.
    Robertson FM, Ogasawara MA, Ye Z et al. (2010) Imaging and Analysis of 3D Tumor Spheroids Enriched for a Cancer Stem Cell Phenotype. J Biomol Screen 15:820-829.

    Answer Id: E15001

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