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CAS:No.168482-84-6
英文名称:Calcein Blue AM
分子式:C21H23NO11
分子量:465.41
储存条件:Powder:-20℃，2 years;Insolvent(母液):-20℃，6 months;-80℃，1 year
纯度:≥95%
说明书下载
Calcein Blue AM是一种细胞可渗透的Calcein Blue。在进入活细胞后，弱荧光Calcein Blue AM被水解为Calcein Blue，其具有类似于DAPI、Hoechst和AMCA的激发/发射最大值。这种与典型的绿色和红色荧光团（如FITC、TMR和Texas red）的特殊光谱分离为多路复用实验提供了额外的选择。由于Calcein Blue AM本身带有荧光，因此可能需要适当的过滤器设置和额外的洗涤步骤来最小化背景荧光。
Flow cytometer（仅供参考）
Excitation 350 nm or 405 nm laser
Emission 450/40 nm filter
Instrument specification(s) Pacific Blue channel
Fluorescence microscope（仅供参考）
Excitation DAPI filter set
Emission DAPI filter set
Recommended plate Black wall/clear bottom
Fluorescence microplate reader（仅供参考）
Excitation 360
Emission 450
Cutoff 420
Recommended plate Black wall/clear bottom
Instrument specification(s) Bottom read mode
Example protocol（仅供参考）
Calcein Blue, AM Stock Solution
Prepare a 2 to 5 mM stock solution of Calcein Blue AM in high-quality, anhydrous DMSO.
If it is necessary to prepare a stock solution, it is recommended to store in aliquots to avoid product failure caused by repeated freezing and thawing.
Note The nonionic detergent Pluronic F-127 can be used to increase the aqueous solubility of AM esters. In the staining buffer, the final Pluronic F-127 concentration should be approximately 0.02%.
Calcein Blue, AM Working Solution
Prepare a Calcein Blue AM working solution of 1 to 10 µM in the buffer of your choice (e.g., Hanks and Hepes buffer). For most cell lines, Calcein Blue AM at the final concentration of 4 to 5 µM is recommended. The exact concentration of indicators required for cell loading must be determined empirically.
Note If your cells contain organic anion-transporters, probenecid (1–2.5 mM) or sulfinpyrazone (0.1–0.25 mM) may be added to the working solution to reduce leakage of the de-esterified indicators.
SAMPLE EXPERIMENTAL PROTOCOL（仅供参考）
Prepare cells for imaging.
Remove the cell culture medium and wash cells once with serum-free buffer to remove any remaining media.
Note Serum in cell culture media may contain esterase activity, which can increase background interference.
Add Calcein Blue AM working solution to the culture.
Incubate cells at 37 ℃ for 30 to 60 minutes.
Replace the dye working solution with HHBS or buffer of your choice (containing an anion transporter inhibitor, such as 1 mM probenecid, if applicable) to remove any excess probes.
Measure the fluorescence intensity using either a fluorescence microscope equipped with a DAPI filter set, a flow cytometer equipped with a 450/40 nm filter (Pacific Blue channel), or a fluorescence plate reader at Ex/Em = 360/450 nm cutoff 420 nm.
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