EdU Cell Proliferation/DNA Synthesis Kit (FACS/Microscopy), Red Fluorescence (BN01108)

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BN01108
  • EdU Cell Proliferation/DNA Synthesis Kit (FACS/Microscopy), Red Fluorescence (BN01108)
  • EdU Cell Proliferation/DNA Synthesis Kit (FACS/Microscopy), Red Fluorescence (BN01108)
  • EdU Cell Proliferation/DNA Synthesis Kit (FACS/Microscopy), Red Fluorescence (BN01108)
€613

Description

ELISA Kit Technical ManualMSDS

EdU Cell Proliferation/DNA Synthesis Kit (FACS/Microscopy), Red Fluorescence

All living cells undergo division cycle, a biological process crucial for proliferation and inheritance. Cell-division cycle is a series of events resulting in two daughter cells containing replicas of DNA from the original DNA molecule. DNA replication occurs in the S phase of the cell cycle and involves de novo synthesis of genomic DNA from its precursors. The ability of monitoring detailed characterization of cell cycle and DNA synthesis in proliferating cells is fundamental in basic, and applied immunologic and oncologic studies. Accurate determination of the effect of biologically active reagents on DNA synthesis and cell cycle is of great importance in anti-cancer drug discovery and basic biology. Assay Genie's EdU DNA Synthesis Monitoring Kit utilizes a novel approach that relies on incorporation of 5-EdU (5-ethynyl-2'deoxyuridine) as nucleoside analog to thymidine into newly synthesized DNA directly in the cell culture. Incorporation of EdU into genomic DNA in S-phase is detected based on a click reaction between the alkyne moiety of EdU and fluorescent azide. Compared to historically used BrdU, click reaction is carried in mild conditions and flow cytometry/fluorescence microscopy can be used for assessment of proliferating cells in the population. Our kit provides sufficient materials for 100 assays based on the protocol below.

Figure: Analysis of newly synthesized DNA in proliferating cells by. (A) HeLa (10 5 cells/ ml) or (B, C) Jurkat (10 6 cells/ ml) cells were incubated with 1X EdU DNA Label for 24 hours in presence and absence of inhibitors of DNA biosynthesis. Red fluorescence reflecting the number of proliferating cells that incorporated the DNA Label in their de novo synthesized DNA was analyzed either by fluorescence microscope (A) or FACS (B, C). (A)Treatment with 10 mM Hydroxyurea (bottom panels) suppressed DNA biosynthesis by blocking DNA replication. Total DNA staining in top and bottom panels clearly confirms that red fluorescence is the result of EdU incorporation during cell proliferation. (B) Jurkat cells incubated with vehicle (red) or in presence of 100 µg/ml Ganciclovir to suppress DNA biosynthesis (green).
Fluorescence measured in FL-2 channel reflects decreased number of proliferating cells treated with Ganciclovir vs. control population. (C) Total DNA content of proliferating cells cultured for 24 h without (green) and with Ganciclovir (red) detected with Total DNA Stain. Fluorescence was measured in FL-1 channel in the linear mode. Ganciclovir arrests proliferating cells in the S phase of the cell cycle. Fixation and Permeabilization followed by detection with Fluorescent Azide and counterstaining with Total DNA Stain was conducted according to the kit protocol.

Key Information Description

Product SKU

BN01108

Size

100 assays

Kit Summary

  • Detection method- Flow Cytometry (Ex/Em 480/(530/590) nm) and Fluorescence Microscopy
  • Sample type- Adherent and suspension cells
  • Species reactivity- Mammalian
  • Applications- This assay provides a convenient and accurate procedure to measure de novo DNA synthesized in biological samples.

Detection Method

Flow Cytometry (Ex/Em 480/(530/590) nm) and Fluorescence Microscopy

Species Reactivity

Mammalian

Applications

This assay provides a convenient and accurate procedure to measure de novo DNA synthesized in biological samples.

Features and Benefits

Simple, fast, does not require lengthy incubation times

Kit Components

  • Wash Buffer (10X)
  • Fixative Solution
  • Permeabilization Buffer (10X)
  • EdU DNA Label (1000X)
  • Copper Reagent (100X)
  • Fluorescent Azide (100X)
  • Reducing Agent (20X)
  • Total DNA Stain (1000X)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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