
EdU Cell Proliferation Kit with Alexa Fluor 555
$230.00 - $730.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: ECK555-500 (for 50-500T)
Cat. No.: ECK555-2k (for 200-2000T)
Description
EdU Cell Proliferation Kit with Alexa Fluor 555 is a reagent kit used for simple, fast, and highly sensitive detection of cell proliferation. It is based on the incorporation of EdU (5-ethynyl-2' -deoxyuridine), a thymidine analog, during DNA synthesis, followed by a click reaction to label EdU with Alexa Fluor 555. This kit can detect individual proliferating cells in cell or tissue samples and quantitatively assess overall cell proliferation in these samples. It is compatible with both cultured cells/tissue samples and tissue sections.
After treatment with this kit, proliferating cells exhibit bright red fluorescence under a fluorescence microscope, which can be detected using various fluorescence detection devices including fluorescence microscopes, laser scanning confocal microscopes, flow cytometers, or fluorescence plate readers. However, flow cytometry or fluorescence plate reader detection is only suitable for cell samples, not tissue sections.
Cell proliferation capacity detection is a fundamental method for evaluating cell activity, genotoxicity, and the effectiveness of anti-tumor drugs. The most accurate method for detecting cell proliferation is direct measurement of DNA synthesis in cells. Traditional methods, such as [3H]thymidine incorporation, have been replaced by antibody-based methods like BrdU (bromo-deoxyuridine) labeling, which, although widely used, has multiple steps, requires BrdU antibodies, and has poor stability. EdU-based methods, on the other hand, do not require antibodies, are easy to perform, and have high detection sensitivity, representing an upgraded replacement for BrdU methods.
Other cell proliferation detection methods based on cell viability, such as MTT assay, WST-1 assay, CCK-8 assay, and chemiluminescence assay, as well as the CFDA SE method based on cell fluorescence tracking, can serve as complementary methods to the EdU method. EdU, also known as 5-ethynyl-2' -deoxyuridine, is a novel thymidine analog that can replace thymidine during DNA synthesis. The alkyne group on EdU can react with fluorescently labeled small molecule azide probes (e.g., Azide Alexa Fluor 488, Azide Alexa Fluor 555, Azide Alexa Fluor 594, Azide Alexa Fluor 647) through a copper-catalyzed click reaction, forming a stable triazole ring. This click reaction allows for the labeling of newly synthesized DNA with the corresponding fluorescent probes, enabling the detection of proliferating cells using appropriate fluorescence detection equipment.
Components
- EdU (10mM)
- Azide 555
- 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 a simple reaction and high detection sensitivity. Based on a straightforward and efficient click reaction, the kit does not require DNA denaturation; only a small amount of azide probe molecules is needed to effectively label the incorporated EdU, allowing for the detection of individual cell proliferation.
- The kit is convenient to use and compatible with various applications. It only requires common paraformaldehyde fixation and Triton X-100 permeabilization to allow the azide probes to effectively enter cells and undergo the click reaction. This process does not affect cell morphology, nor does it interfere with antibody-based immunofluorescence and immunohistochemistry detection. Furthermore, it does not interfere with DNA fluorescence staining (such as PI staining for cell cycle analysis, or DAPI or Hoechst staining for nuclear detection). In contrast, the BrdU method requires denaturation of double-stranded DNA (e.g., acid denaturation, heat denaturation, or DNase digestion) to allow large BrdU antibodies to enter cells and bind to BrdU on DNA. This denaturation process 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 takes at least 4 hours, our proprietary EdU method used in this kit for detecting newly synthesized DNA only requires 1.5-2 hours, significantly reducing the time needed. Additionally, the kit provides Hoechst 33342 for staining cell nuclei, facilitating 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 555 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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