Invitrogen™ Click-iT™ EdU Imaging Kit with Alexa Fluor™ 488, 594, and 647 Azides

The Click-iT EdU Imaging Kit is a trial version of our superior alternative to traditional proliferation assays optimized for fluorescence microscopy applications. In this assay the modified thymidine analogue EdU is efficiently incorporated into newly synthesized DNA and fluorescently labeled with a bright, photostable Alexa Fluor dye in a fast, highly specific click reaction. This fluorescent labeling of proliferating cells is accurate and compatible with antibody methods due to the mild click protocol. This trial-size kit provides sufficient EdU to label up to 6 to 30 coverslips and enough Alexa Fluor 488, 594, and 647 azide to detect new DNA synthesis on 2 to 10 coverslips with each fluorophore. The most accurate proliferation-detection methods are based on the incorporation and measurement of nucleoside analogues in newly synthesized DNA, with bromodeoxyuridine (BrdU) a commonly used analogue. BrdU-labeled DNA is quantitated using anti-BrdU antibodies following DNA denaturation by harsh methods (HCl, heat, or enzymes) to expose the BrdU molecules. This step is time consuming and difficult to perform consistently. The harsh treatment can also adversely effect sample integrity and quality, which makes co-staining with other antibodies challenging.

• Simple: works the first time, every time, in less time
• Efficient: no denaturation steps or harsh treatment required
• Content-rich results: better preservation of cell morphology, antigen structure, and DNA integrity
• Consistent: not dependent on variable antibody lots for detection
• Trial version

Superior Proliferation Methodology
The Click-iT EdU Imaging Kit provides a superior alternative to BrdU assays for measuring cell proliferation. EdU (5-ethynyl-2'-deoxyuridine) is a nucleoside analog of thymidine and is incorporated into DNA during active DNA synthesis. With Click-iT EdU, mild fixation and detergent permeabilization is sufficient for the small molecule-based Click-iT EdU detection reagent to gain access to the DNA. As a consequence, the Click-iT EdU Imaging Kit is not only easy to use, but more accurate and compatible with cell cycle analysis and other intracellular or extracellular targets for truly content-rich results.

The Click-iT assay can be used on cells in culture or in vivo following EdU administration by feeding or injection methods. The Click-iTassay can be used with BrdU in dual pulse experiments by using the anti-BrdU (clone MoBu-1) antibody, which does not cross react with EdU. The Click-iTtechnology is compatible with immunohistochemical, immunocytochemical, and fluorescent dyes that are fixation tolerant or designed for fixed cell labeling.

Cell Analysis, Cell Proliferation, Cell Proliferation with Click Chemistry, Cell Viability, Proliferation and Function, Cellular Imaging, Click Chemistries, Immunofluorescence (IF), Immunofluorescence Staining and Detection, Labeling Chemistry