Key features and details
- Assay type: Cell-based
- Detection method: Fluorescent
- Platform: Flow cytometer, Fluorescence microscope
- Sample type: Adherent cells, Suspension cells
Product nameEdU Assay / EdU Staining Proliferation Kit (iFluor 488)
Sample typeAdherent cells, Suspension cells
EdU Assay / EdU Staining Proliferation Kit (iFluor 488) ab219801 provides a sensitive and robust method to detect and quantify cell proliferation in live mammalian cells using flow cytometry or fluorescence microscopy. The iFluor 488 dye (Ex/Em: 491/520 nm) has spectral properties almost identical to those of FITC and alternative green 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 488).
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 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
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
PlatformFlow cytometer, Fluorescence microscope
Storage instructionsStore at -20°C. Please refer to protocols.
Components 50 tests 50 tests 10X Permeabilization Buffer 1 x 25ml 1 x 25ml Copper Sulfate (100 mM) 1 x 1ml 1 x 1ml Dimethylsulfoxide (DMSO) 1 x 4.25ml 1 x 4.25ml EdU 1 x 10mg 1 x 10mg Fixative (40% formaldehyde solution) 1 x 5ml 1 x 5ml iFluor 488 azide dye (500 μM) 1 x 130µl 1 x 130µl Sodium Ascorbate 1 x 400mg 1 x 400mg
Dot plot of EdU-488 staining (Y-axis, 488) vs FSC. 106 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were incubated with the stated concentrations of EdU for 3 hours. Control cells (next image) were incubated with media only. Images were acquired on an Accuri C6 Cytometer (BD Biosciences) with cells excited using a 488 nm laser and data analyzed using FlowJo (v10). The percentage of gated cells (EdU positive) is highlighted.
EdU staining of proliferating cells. HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (4 x 104 cells/well in 96 plate) were incubated with 20 μM EdU for 3 hours. Cells were analyzed using a TCS SP8 confocal microscope (Leica-Microsystems). DNA (blue) was staining with Hoechst 33342 ab145597. Green cells show EdU/Hoechst-positive cells.
Dot plot of EdU-488 staining (Y-axis, 488) vs FSC. 106 HeLa (Human epithelial cell line from cervix adenocarcinoma) cells were incubated with the stated concentrations of EdU for 3 hours. This image shows control cells, incubated with media only. Images were acquired on an Accuri C6 Cytometer (BD Biosciences) with cells excited using a 488 nm laser and data analyzed using FlowJo (v10). The percentage of gated cells (EdU positive) is highlighted.
EdU staining of proliferating cells. HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (4 x 104 cells/well in 96 plate) were incubated with 10 μM EdU for 3 hours. Cells were analyzed using a TCS SP8 confocal microscope (Leica-Microsystems). DNA (blue) was staining with Hoechst 33342 ab145597. Green cells show EdU/Hoechst-positive cells.
ab219801 has been referenced in 10 publications.
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- Dilasser F et al. Essential role of smooth muscle Rac1 in severe asthma-associated airway remodelling. Thorax 76:326-334 (2021). PubMed: 33542087
- Xie JW et al. Alpha defensin-1 attenuates surgically induced osteoarthritis in association with promoting M1 to M2 macrophage polarization. Osteoarthritis Cartilage 29:1048-1059 (2021). PubMed: 33892137
- Guo QQ et al. RNA-Binding Protein MSI2 Binds to miR-301a-3p and Facilitates Its Distribution in Mitochondria of Endothelial Cells. Front Mol Biosci 7:609828 (2020). PubMed: 33553241
- Mal A et al. EpCAM-Mediated Cellular Plasticity Promotes Radiation Resistance and Metastasis in Breast Cancer. Front Cell Dev Biol 8:597673 (2020). PubMed: 33490064
- Shao B et al. RP11-284F21.9 promotes oral squamous cell carcinoma development via the miR-383-5p/MAL2 axis. J Oral Pathol Med 49:21-29 (2020). PubMed: 31397491
- Wang Z et al. Long Noncoding RNA DIO3OS Hinders Cell Malignant Behaviors of Hepatocellular Carcinoma Cells Through the microRNA-328/Hhip Axis. Cancer Manag Res 12:3903-3914 (2020). PubMed: 32547226
- Zhou C et al. Bifidobacterium longum alleviates irritable bowel syndrome-related visceral hypersensitivity and microbiota dysbiosis via Paneth cell regulation. Gut Microbes 12:1782156 (2020). PubMed: 32584650
- Li D et al. RP11-284F21.9 promotes lung carcinoma proliferation and invasion via the regulation of miR-627-3p/CCAR1. Oncol Rep 44:1638-1648 (2020). PubMed: 32945522
- Li J et al. Liraglutide protects renal mesangial cells against hyperglycemia-mediated mitochondrial apoptosis by activating the ERK-Yap signaling pathway and upregulating Sirt3 expression. Mol Med Rep 19:2849-2860 (2019). PubMed: 30816450