Measurement of Poly(ADP-ribose) Level with Enhanced Slot Blot Assay with Crosslinking
Abstract
:1. Introduction
2. Results
2.1. Optimization of Slot Blot Assay of PAR Using Crosslinking
2.2. Effects of PAR Length on Detection with Slot Blot Assay of PAR
2.3. Detection of PAR Level Changes with Slot Blot Assay in Cultured Cells
2.4. PAR Levels in Mouse PBMCs Detected with Ex Vivo PARP Assay
3. Discussion
4. Materials and Methods
4.1. Preparation of PAR Using Recombinant PARP-1
4.2. Slot Bot Assay of PAR
4.3. Ex Vivo Assay of PARP Activity
4.4. Extraction of PBMC from Mice
4.5. UV Crosslinking
4.6. Chemical Crosslinking
4.7. Culture of Mouse Wild-Type ES Cells
4.8. Statistical Analysis
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Kudo, Y.; Sasaki, Y.; Onodera, T.; Hashimoto, J.; Nozaki, T.; Tamura, K.; Watanabe, M.; Masutani, M. Measurement of Poly(ADP-ribose) Level with Enhanced Slot Blot Assay with Crosslinking. Challenges 2018, 9, 27. https://doi.org/10.3390/challe9020027
Kudo Y, Sasaki Y, Onodera T, Hashimoto J, Nozaki T, Tamura K, Watanabe M, Masutani M. Measurement of Poly(ADP-ribose) Level with Enhanced Slot Blot Assay with Crosslinking. Challenges. 2018; 9(2):27. https://doi.org/10.3390/challe9020027
Chicago/Turabian StyleKudo, Yuko, Yuka Sasaki, Takae Onodera, Jun Hashimoto, Tadashige Nozaki, Kenji Tamura, Masatoshi Watanabe, and Mitsuko Masutani. 2018. "Measurement of Poly(ADP-ribose) Level with Enhanced Slot Blot Assay with Crosslinking" Challenges 9, no. 2: 27. https://doi.org/10.3390/challe9020027