Invitrogen™ Click-iT™ EdU Alexa Fluor™ 647 HCS Assay

Description

• The Click-iT EdU Alexa Fluor 647 HCS Assay is a superior alternative to traditional proliferation assays that is optimized for high-content imaging 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
• 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 Less hazardous Superior Proliferation Methodology The Click-iT EdU Alexa Fluor 647 HCS Assay 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 HCS assay 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 administration of EdU by feeding or injection methods
• The Click-iT assay can be used with BrdU in dual pulse experiments by using the ant-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 Cycle, Cell Proliferation, Cell Viability, Proliferation and Function, Cellular Imaging, DNA Replication and Repair, High-Content Screening (HCS) Order Info Shipping Condition: Room temperature