EdU Cell Proliferation Kit with Alexa Fluor 647
$230.00 - $730.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: ECK647-500 (for 50-500T)
Cat. No.: ECK647-2k (for 200-2000T)
Description
EdU Cell Proliferation Kit with Alexa Fluor 647 is based on the incorporation of 5-ethynyl-2'-deoxyuridine (EdU), a thymidine analogue, during DNA synthesis, followed by a click reaction to label EdU with Alexa Fluor 647. This kit enables simple, rapid, and highly sensitive detection of cell proliferation.
It can detect individual proliferating cells in cell or tissue samples, as well as quantitatively analyze the overall cell proliferation in these samples. The kit is applicable to cultured cells, tissue samples, and tissue sections.
After treatment with this kit, proliferating cells exhibit strong near-infrared fluorescence under laser confocal microscopy. While this fluorescence may not be visible to the naked eye upon excitation, it can be detected using various imaging systems such as laser confocal microscopy or flow cytometry equipped with corresponding excitation and emission detection modules. Additionally, it can be used for High-Content Screening (HCS). However, flow cytometry or fluorescence enzyme labeling detection is only suitable for cell samples and not tissue sections.
Detecting cell proliferation is fundamental for assessing cell viability, genotoxicity, and the efficacy of anti-tumor drugs. The most accurate method for detecting cell proliferation is directly measuring DNA synthesis in cells. Initially, radioactive nucleotide incorporation methods, such as [3H]thymidine incorporation, were widely used for this purpose. However, due to radioactive contamination and difficulties in single-cell detection, these methods have been largely replaced by BrdU assays based on antibody detection. Nevertheless, BrdU assays are cumbersome, require BrdU antibodies, and exhibit various influencing factors, resulting in poor stability. Moreover, interference may occur when BrdU assays are combined with other antibody-based detection methods. The EdU method, based on EdU incorporation and subsequent click reactions, does not require antibodies, is easy to perform, and offers high detection sensitivity. It represents a new approach that upgrades and replaces BrdU assays.
Methods such as MTT, WST-1, CCK-8, and chemiluminescence assay are all cell proliferation detection methods based on cell activity, capable of detecting the overall proliferation effect of cells. However, they cannot detect individual proliferating cells. Although these methods do not detect DNA synthesis, they are widely used as alternatives to [3H]thymidine incorporation assays. CFDA SE, based on cell fluorescence tracking principles, can detect individual proliferating cells. However, due to fluorescence halving with each cell division, it is challenging to distinguish cells with halved fluorescence under a fluorescence microscope, and its detection sensitivity is not very high. It is usually only suitable for flow cytometry. In scientific research, these methods based on cell activity or CFDA SE can serve as supplementary detection methods for the EdU method.
EdU, also known as 5-ethynyl-2'-deoxyuridine, is a novel analogue of thymidine. EdU can replace thymidine during DNA synthesis, incorporating into newly synthesized DNA. Moreover, the alkyne group on EdU can react with fluorescently labeled small molecule azide probes through a copper-catalyzed click reaction, forming stable triazole rings. This reaction, known as the click reaction, is extremely rapid. Through click reactions, newly synthesized DNA can be labeled with corresponding fluorescent probes, enabling detection of proliferating cells using appropriate fluorescence detection equipment.
Components
- EdU (10mM)
- Azide 647
- Click Reaction Buffer
- CuSO4
- Click Additive
- Hoechst 33342 (1000X)
Specification
The small package of the kit can detect 50 samples when used for cell cultures (6-well plate), with a reaction volume of 500 μl of Click reaction solution per sample. If used for detection in a 96-well plate, it can detect 500 samples, with a detection system volume of 50 μl of Click reaction solution per sample. For detection in 12-well, 24-well, 48-well, or 384-well plates, it can detect 125, 250, 350, and 1250 samples respectively, with recommended volumes of 200 μl, 100 μl, 70 μl, and 20 μl of Click reaction solution per sample.
For flow cytometry detection, the small package can detect 50 samples, with an ideal cell quantity of 10-100 million cells per sample, and a reaction volume of 500 μl of Click reaction solution per sample.
For detection in frozen or paraffin-embedded sections, the small package can detect 125-250 samples, with a reaction volume of 100-200 μl of Click reaction solution per sample.
The large package can detect four times the number of samples as the small package.
Features
- This kit offers simple reaction procedures and high detection sensitivity. Based on a straightforward and efficient Click reaction, it efficiently labels incorporated EdU without the need for DNA denaturation, requiring only a small amount of small molecule azide probes to effectively mark the EdU incorporation, enabling the detection of individual cell proliferation.
- The kit is convenient to use and exhibits good compatibility. With only common formaldehyde fixation and Triton X-100 permeabilization, the azide probes can effectively enter cells and undergo Click reaction, without affecting cell morphology, antibody-based immunofluorescence, immunohistochemistry, or DNA fluorescence staining (such as PI staining for cell cycle detection, DAPI or Hoechst dyes for nuclear staining). In contrast, the BrdU method requires denaturation of double-stranded DNA (such as acid denaturation, heat denaturation, or DNase digestion) to allow large BrdU antibodies to enter cells and bind to BrdU on DNA, which may affect cell morphology and subsequent immunofluorescence, immunohistochemistry, and DNA fluorescence staining.
- This kit offers rapid detection, making both qualitative and quantitative analyses highly convenient. Compared to the BrdU method, which typically requires at least 4 hours, our proprietary EdU method employed by this kit for detecting newly synthesized DNA takes only 1.5-2 hours, significantly reducing the time required. Additionally, the kit provides Hoechst 33342 for staining cell nuclei, facilitating the observation of all cell nuclei. Qualitative and quantitative analyses can be performed using fluorescence microscopy or flow cytometry.
Storage
Store at -20°C, valid for one year. Azide 647 and Hoechst 33342 should be stored protected from light.
Notes
- After preparing the Click Additive into a solution, please ensure appropriate aliquoting. If white precipitates appear after dissolution, invert the solution several times until fully dissolved before use. If the solution turns brown, it indicates that the active component of this ingredient has deteriorated, and the solution should be discarded.
- If hydroxyurea is needed as a control, it can be ordered from us.
- If more EdU is required for animal experiments, it can also be ordered from us.
- Due to the copper ion catalysis required for the click reaction, please note the following compatibility issues and solutions: This product is fully compatible with organic dyes such as the Alexa Fluor® series and fluorescein (FITC), Allophycocyanin (APC), and APCE-tandem dyes. For Qdot® nanocrystal probes, Horseradish peroxidase (HRP), R-phycoerythrin (R-PE), and R-PE-tandem dyes such as Alexa Fluor® 680-R-PE, reactions and detection should be performed after the click reaction is completed. This product will affect the fluorescence of GFP, RFP, mCherry, and similar fluorescent proteins. For fluorescent proteins such as Green Fluorescent Protein (GFP), TC-FlAsH™, and TC-ReAsH™ reagents, reactions and detection should be performed before the click reaction. Since Phalloidin is not compatible with the click reaction, it is recommended to use Tubulin-Tracker Red for microtubule detection.
- This product is for scientific research by professionals only and should not be used for clinical diagnosis or treatment, food or drug purposes, or stored in ordinary residences.
- For your safety and health, wear laboratory clothing and disposable gloves during operation.
Only for research and not intended for treatment of humans or animals
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