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Контроль синтеза ДНК (Пролиферация клеток)

... on DNA - DNA synthesis monitoring (Cell proliferation)

Cell proliferation is characterized by de novo DNA synthesis during the S-Phase of the cell cycle. The traditional approach for monitoring DNA synthesis and thus cell proliferation relies on the incorporation of 5-Brom-deoxyuridine (5-BrdU) into nascent DNA strands by cellular DNA polymerases. This method is hampered by the cell morphology destructive detection via a BrdU specific antibody that requires harsh cell permeabilization conditions and nucleic acid denaturation procedures preventing subsequent analysis of other cellular targets for multiparametric analysis.

This drawback can be bypassed by the incorporation of ethynyl-labeled deoxyuridine (5-EdU[1,2,3,4]) or ethynyl-labeled deoxycytidine (5-EdC[5]) that are incorporated into nascent DNA strands of proliferating cells instead of their natural counterparts (thymidine or cytidine, respectively). The ethynyl-functionalized DNA is subsequently detected by covalent conjugation of small-sized fluorescent Azides or Azides of Biotin via Cu(I)-catalyzed Click Chemistry (Fig. 1).

5-EdU & 5-EdC-based nucleic acid labeling

  • is non-destructive and therefore suitable for mulitparametric analysis (no harsh permeabilization & nucleic acid denaturation required).
  • provides a significantly faster detection procedure (∼2 h instead of 4 h +).
  • is compatible with flow cytometry & microscopy[1,4,5,6]
  • is compatible with several cell lines and organisms (selected cell lines & organisms analyzed with 5-EdU)
 
Click Chemistry
Figure 1 Workflow of DNA synthesis monitoring with 5-EdU. The labeling procedure with 5-EdC is similar.
 

НазваниеКодЦена Количество
10 mg pack CLK-N003-10 16427.84 руб. 10 mg



НазваниеКодЦена Количество
100 mg pack CLK-N001-100 24305.94 руб. 100 mg
25 mg pack CLK-N001-25 8377.53 руб. 25 mg
5 g pack CLK-N001-5000 405099 руб. 5 g
500 mg pack CLK-N001-500 97223.76 руб. 500 mg




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