1. Simple: no antigen-antibody reaction is required; the detection method based on small molecule chemical reaction is simple and efficient, and the reaction takes only a few minutes
2. Sensitive: no antibodies required, only 1/500 of BrdU antibody needed for the detection of dye, easy to spread, even a single proliferating cell can be accurately detected
3. Fast: it takes only 5 hours to complete the entire inspection cycle by eliminating the complicated steps of antigen-antibody reaction
4. Accurate: no need for DNA denaturation (acid hydrolysis, pyrolysis, enzymatic hydrolysis, etc.), so as to effectively avoid sample damage and ensure clear and intact nucleus margin
5. Compatible: It has almost no damage to the sample, easier to label with multiple antibodies or fluorescent proteins. It can simultaneously detect other traits of cells.
6. Multiple Options: a variety of dyes are optional, suitable for a variety of animal cell detection, and also suitable for plant cells

Multiple Dye Options

RiboBio EdU products provide a variety types of dye for multiple detection purposes. It maximizes the applicable scope of third-party dyeing, suits the needs of testing instruments, and helps solve the experimental problems.

Excitation spectrum and emission spectrum of three dyes (dashed line: excitation spectrum; solid line: emission spectrum)
Note: The chemical reaction of Apollo staining will inactivate GFP, EGFP, and R-PE.

Three dyes were respectively used in high-content screening platform (BD pathway 855) to detect HeLa cells (40X) that had been incubated with 50uM EdU for 2 hours. The Apollo®643 dye detection result was in gray-scale value, and the picture was rendered in red.

Simple: no antigen-antibody reaction is required,  the detection method based on small molecule chemical reaction is simple and efficient, and the reaction takes only a few minutes

Figure 1. Schematic comparision of BrdU and EduU detection methods

Sensitive: no antibodies is required, only 1/500 of BrdU antibody needed for the detection of dye, easy to spread, even a single proliferating cell can be accurately detected

Cell proliferation of A549 cells incubated for 30 minutes

Cell proliferation of Hela cells incubated for 1 hour

Cell proliferation of 3T3 cells incubated for 6 hours

Fast: it takes only 2.5 hours to complete the entire inspection cycle by eliminating the complicated steps of antigen-antibody reaction

10X

20X

40XFigure 7.  Cell proliferation assay of A549 cells incubated for 2 hours (10X, 20X, 40X)

Accurate: no need for DNA denaturation (acid hydrolysis, pyrolysis, enzymatic hydrolysis, etc.) to effectively avoid sample damage and ensure clear and intact nucleus margin

40X

Figure 8. BrdU requires DNA denaturation to bind to antibodies, resulting in BrdU, Hoechst staining and blurred edges; but EdU edges are clear and complete, indicating that it’s more sensitive and accurate

Compatible: It has almost no damage to the sample, easier to label with multiple antibodies or fluorescent proteins. It can simultaneously detect other traits of cells.

The EdU cell proliferation assay does not require drastic DNA denaturation, it only needs the gentle cell immobilization and permeabilization to better protect the cell morphology, DNA structure and intracellular antigen recognition sites. It’s more suitable to multi-label with other dyes or protein labeling (eg: P65 antibody), directly access multi-dimensional cell features at the cellular leve, it’s also more suitable for the in-depth research on cell function.

Analysis of the effects of drugs on cell proliferation and NF-KB signaling pathway in combination with P65 antibody and EdU

Recommended for a variety of animal/plant cell tests

The simple, sensitive, fast, and accurate EdU proliferation assay is applicable to a variety of cell lines.

The wall of plant cells is relatively thick and plays a protective role for plants. The antibody detection method usually entails the digestion of the cell wall, which may lead to impurity interference and even damage the cells, and it can reduce the reliability of detection. Both EdU and Apollo® dye molecules are small and can easily penetrate the cell wall, so the plant growth, development, differentiation can be accurately detected without digesting cell walls, or DNA denaturation and antigen-antibody reaction.

Detection of plant cell growth and differentiation by EdU (Kotogány E, et al. Plant Methods. 2010)