EdU Assay / EdU Staining Proliferation Kit (iFluor 647)

EdU Assay / EdU Staining Proliferation Kit (iFluor 647) ab222421 provides a sensitive and robust method to detect and quantify cell proliferation in live mammalian cells using flow cytometry or fluorescence microscopy. The iFluor 647 dye (Ex/Em: 649/664 nm) has spectral properties almost identical to those of Cy5^®and alternative red fluorophores.

EdU staining protocol summary (wash cells between each step):
- add EdU solution to cells to be stained
- incubate cells for 2-4 hrs under optimal growth conditions
- add fixative solution and incubate for 15 min
- add permeabilization buffer and incubate for 15/20 min
- add reaction mix to fluorescently label EdU and incubate for 30 min
- analyze with flow cytometer / fluorescence microscope

EdU staining can also be combined with antibody staining or cell staining with other fluorescent dyes.

This kit provides enough reagents to perform 50 flow cytometry tests or 50 microscopy tests (for 18 x 18 mm coverslips) or 200 microscopy tests (adapted for 96-well plate format).

Previously called EdU Proliferation Assay Kit (iFluor 647).The most accurate method to measure DNA proliferation is by directly measuring DNA synthesis. The most common method for this uses antibody-based detection of the nucleoside analog bromo-deoxyuridine (BrdU). EdU (5-ethynyl-2’-deoxyuridine), a thymidine analog that is an alternative to BrdU, is also used in DNA proliferation assays that are simpler and faster than the BrdU assay. NB: EdU is also available as free molecule as EdU, DNA synthesis monitoring probe ab146186 (EdU).In EdU staining, EdU is incorporated into newly synthesized DNA by cells within a sample. A fluorescent azide, such as iFluor-488, is then added. The fluorescent azide is small enough to diffuse freely through native tissues and DNA, and it covalently cross-links to the EdU in a 'click' chemistry reaction. The main advantages of EdU staining over using BrdU are: - no harsh DNA hydrolysis / DNA denaturing step is required with EdU staining (unlike in the BrdU assay where it is used to give the BrdU antibody access to BrdU within the DNA) - EdU staining is faster, and has less steps, than BrdU staining

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