E-Click EdU Cell Proliferation Imaging Assay Kit (Green, FITC)
E-Click EdU Cell Proliferation Imaging Assay Kit (Green, FITC)
Introduction
Elabscience® E-Click EdU Cell Proliferation Imaging Assay Kit is easy to operate and has high sensitivity. It is suitable for the proliferation assay of cell slides and smears, and the results can be observed by flow fluorescence microscope.
Product Details
Properties
| Application | Cell Proliferation |
| Detection method | Fluorometric Method |
| Sample type | Cell Slides, Cell Smears |
| Assay time | 2 hours |
| Detection instrument | Fluorescence Microscope |
| Dye Type | FITC |
| Ex/Em (nm) | 490/520 |
| Filter set | FITC |
| Other reagents required | PBS (pH7.2~7.6)(E-IR-R187), PBS buffer (with 3%BSA) (PH7.2 ~ 7.4), PBS (with 0.3% Triton X-100) (pH7.2~7.6), 4% Polyformaldehyde (dissolved in PBS)(E-IR-R113), Deionized water. |
| Storage | -20°C, shading light |
| Shipping | Ice bag |
| Expiration date | 12 months |
Detection Principle
Cell proliferation assays are widely used in the evaluation of cell viability, genotoxicity, and the effect of antitumor drugs. Direct detection of DNA synthesis in cells is considered to be the most accurate method for detecting cell proliferation. The initial widely used method for detecting DNA synthesis in cells was the radiolabeled nucleoside incorporation method, but this method was greatly limited due to radioactive contamination and the difficulty of single-cell detection, and was gradually replaced by the BrdU method based on antibody detection. The BrdU method has many steps and requires the use of BrdU antibody, which has many influencing factors and poor stability.
EdU method is based on EdU incorporation and subsequent click reaction, without the use of antibodies, convenient operation and high detection sensitivity. It is a new method upgraded on the basis of BrdU method and will gradually replace BrdU method. EdU (5-ethynyl-2-deoxyuridine), is a thymidine analog, EdU can replace thymidine in the process of DNA synthesis to incorporate into new in synthetic DNA. On the other hand, the acetylene group on EdU can react with fluorescently labeled small molecule azide probes (such as FITC Azide, Elab Fluor® 488 Azide, Elab Fluor® 594 Azide, Elab Fluor® 647 Azide) through the catalysis of monovalent copper ions to form a stable triazole ring. This reaction is very rapid and is called the click reaction. Through the click reaction, the newly synthesized DNA is labeled with the corresponding fluorescent probe, so that the proliferating cells can be detected using the appropriate fluorescent detection equipment.
Storage
Store at -20°C for 12 months. EdU (10 mM) needs to be stored in aliquots (50 μL/vial is recommended or aliquot into smaller quantities according to experimental needs) for the first use.
Notes
- This kit is for research use only.
- Please take safety precautions and follow the procedures of laboratory reagent operation.
- The labeling concentration of EdU should be optimized according to the cell type used. It is recommended to do a preliminary experiment to explore the optimal concentration of EdU and 10 μM EdU can be used as initial exploratory concentration.
- Since the EdU labeling reaction is carried out in the cells and detected by fluorescence microscope, please ensure that the cells are completely fixed and permeabilized before EdU labeling. If the room temperature is too low such as in winter, it is recommended to extend the fixation time appropriately or fix it overnight at 4°C.
- Aliquot the Click Additive Solution and store at -20ºC. If white substance is precipitated before use, please turn it upside down several times and use it only after it has completely dissolved. If the color of the Click Additive Solution turns brown, indicates that the reagent has expired, please discard it.
- Copper ions will affect the fluorescence of GFP, RFP, mCherry and other fluorescent proteins, so this kit is not suitable for cells with GFP, RFP, mCherry and other fluorescence.