Alexa Fluor ® 488 Phalloidin

品牌： CST

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参数

实验应用：

IF-F/IF-IC

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Product Usage Information

To make a stock concentration of 6.6 μM (20X), reconstitute the lyophilized material in 1.5 ml pure methanol or 150ul of anhydrous DMSO. Fix cells for 15 minutes using fresh, methanol-free 4% formaldehyde, then rinse three times in PBS for 5 minutes each. Once fixed, please follow CST protocol for immunostaining. Following incubation of the primary and secondary antibodies, Alexa Fluor® 488 Phalloidin can be diluted 1:20 (5 μl per 100 μl assay volume) in PBS and added to the cells. Allow to incubate for 15 minutes at room temperature, then rinse once with PBS. Coverslip slides with ProLong® Gold Antifade Reagent #9071 and examine specimen using appropriate excitation wavelength.

应用

实验应用

IF-F/IF-IC

背景

Actin, a ubiquitous eukaryotic protein, is the major component of the cytoskeleton. At least six isoforms are known in mammals. Nonmuscle β- and γ-actin, also known as cytoplasmic actin, are predominantly expressed in nonmuscle cells, controlling cell structure and motility (1). Actin exists mainly as a fibrous polymer, F-actin. In response to cytoskeletal reorganizing signals during processes such as cytokinesis, endocytosis, or stress, cofilin promotes fragmentation and depolymerization of F-actin, resulting in an increase in the monomeric globular form, G-actin (2). Phalloidin is a natually occuring toxic bicyclic peptide found in the deathcap toadstool, Amanita phalloides, that rapidly binds to F-actin with strong affinity (3). 1.Herman, I.M. (1993) Curr Opin Cell Biol 5, 48-55. 2.Condeelis, J. (2001) Trends Cell Biol 11, 288-93. 3.Lengsfeld, A.M. et al. (1974) Proc Natl Acad Sci U S A 71, 2803-7.

制备和贮存

保存方式

This material is provided as lyophilized solid that is stable for 2 years at -20°C, desiccated and protected from light. Phalloidin conjugates should be reconstituted in pure methanol or anhydrous DMSO to make stock solutions, please refer to the directions for use for details. Once reconstituted in pure methanol or anhydrous DMSO, stock solutions are stable for 1 year at -20°C, desiccated and protected from light. Stability in aqueous solutions is low and the conjugate should only be in the presence of an aqueous solution during incubation with cells.

参考图片

Confocal immunofluorescent analysis of HeLa cells using Alexa Fluor® 488 Phalloidin (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).共聚焦免疫荧光分析HeLa细胞，使用Alexa Fluor® 488 Phalloidin（绿色）。蓝色假色= DRAQ5® #4084（DNA荧光染料）。

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货号：

8878S

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