Alexa Fluor? 647 Phalloidin

• 产品编号：CST-8940S      品牌：CST       原厂货号：8940S
• 产品分类：生化试剂 > 染料/染色剂/指示剂/底物 > 细胞染料 > 其他
• 应用分类：

描述：

对于6.6 μM (20X)浓度的储存液，将冻干粉溶于1.5 ml纯甲醇中。固定细胞，用新鲜无甲醇的4%甲醛固定15分钟，然后用PBS清洗3次，每次5分钟。一旦固定，根据CST公司提供的方案进行免疫染色。加入一抗和二抗进行孵育，Alexa Fluor® 647鬼笔环肽用PBS 1：20稀释（100μl体积中加入5μl）后加入细胞中。在室温下孵育15分钟后，用PBS清洗一次。盖玻片上滴加ProLong® Gold 抗淬灭试剂，用合适的激发波长检测样品。肌动蛋白是一种普遍存在的真核细胞的蛋白，是细胞骨架的主要成分。在哺乳动物中至少有六种亚型。非肌肉β -和γ-肌动蛋白，也被称为细胞质肌动蛋白，主要在非肌肉细胞中表达，控制细胞的结构和运动（1）。肌动蛋白主要以纤维多聚体形式存在，称为丝状肌动蛋白。针对胞质分裂、胞吞作用和应激过程中的细胞骨架的重组信号，人肌动蛋白素能促进丝状肌动蛋白的分裂和解聚，使单体球状形式的肌动蛋白增加，也称为球形肌动蛋白（2）。鬼笔环肽是一种自然出现的有毒的二环肽，存在于剧毒毒伞的子实体菌、鬼笔鹅膏中，对丝状肌动蛋白有很强的亲和力（3）。

原厂资料：

Directions for Use

To make a stock concentration of 6.6 μM (20X), reconstitute the lyophilized material in 1.5 ml pure methanol. 
 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® 647 Phalloidin can be diluted 1:20 (5 μl per 100 μl assay volume) in PBS and added to cells. Allow to incubate for 15 minutes at room temperature, then rinse once with PBS. Coverslip slides with ProLong® Gold antifade reagent and examine specimen using appropriate excitation wavelength.

Background

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).

注意事项：

Storage: This material is provided as lyophilized solid that is
stable for 1 year at -20°C, desiccated and protected from light.
Stock solutions are stable for 1 year at -20ºC, desiccated and
protected from light. Phalloidin conjugates should be recon-
stituted in pure methanol to make stock solutions, please refer
to the directions for use for details. Once reconstituted in pure
methanol, stock solutions are stable for 1 year at -20°C, desic-
cated 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