产品
编 号:F368768
分子式:C17H19N3
分子量:265.35
产品类型
结构图
CAS No: 494-38-2
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产品详情
生物活性:
Acridine Orange base is a cell-permeable fluorescent dye that stains organisms (bacteria, parasites, viruses, etc.) bright orange and, when used under appropriate conditions (pH=3.5, Ex=460 nm), distinguishes human cells in green for detection by fluorescence microscopy. Acridine Orange base fluoresces green when bound to dsDNA (Ex=488, Em=520-524) and red when bound to ssDNA (Ex=457, Em=630-644) or ssRNA (Ex=457, Em=630-644), also can be used in cell cycle assays.
体外研究:
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).1. Differential staining of DNA and RNA of unfixed cells:(1) Set up the flow cytometer with excitation at 488 nm, using emission filters and a dichroic mirror that discriminate green fluorescence (measured at 515-545 nm) and red luminescence (measured preferably above 640 or 650 nm).(2) Transfer a 0.2-mL aliquot of the original cell suspension to a small glass or plastic tube (e.g., 2- or 5-mL volume). Chill on ice.(3) Gently add 0.4 mL ice-cold cell permeabilizing solution. Wait 15 s, keeping cells on ice.(4) Gently add 1.2 mL ice-cold Acridine Orange base staining solution. Keep cells on ice in the dark.(5) Measure and record cell fluorescence in the flow cytometer during the 2 to 10 min after addition of Acridine Orange base staining solution.2. Differential staining of fixed cells:(1a) For cells in suspension culture or hematologic samples: Rinse cells once with ice-cold PBS and suspend in ice-cold PBS at ~106 cells/mL.(1b) For cells attached to tissue culture plates: Collect cells from flasks or petri plates by trypsinization, pool the trypsinized cells with cells floating in the medium (mostly detached mitotic and dead cells), and rinse once with medium containing serum to inactivate the trypsin. Suspend cells in ice-cold PBS at ~106 cells/mL.(1c) For cells isolated from solid tumors: Rinse cells free of any enzyme used for cell dissociation and suspend in ice-cold PBS at ~106 cells/mL.(2) With a Pasteur pipet transfer 1 mL cell suspension to a 15-mL conical glass tube containing 10 mL ice-cold 70% ethanol. Fix cells ≥2 h on ice.(3) Centrifuge tubes 5 min at 300 × g, 4℃. Remove all ethanol, rinse cells once with ice-cold PBS, and suspend in ice-cold PBS at a density of Acridine Orange base 相关抗体: