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12935
DyLight™ 488 Phalloidin
Buffers & Dyes
Cellular Dyes

DyLight™ 488 Phalloidin #12935

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  1. IF
Immunofluorescence Image 1: DyLight™ 488 Phalloidin

Confocal immunofluorescent analysis of HeLa cells using Cox IV (4D11-B3-E8) Mouse mAb #11967 (red). Actin filaments were labeled with DyLight™ 488 Phalloidin (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Product Usage Information

To make a 40X stock, 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 Cell Signaling Technology protocol for immunofluorescence. Following incubation of the primary and secondary antibodies, DyLight™ 488 Phalloidin can be diluted 1:40 (2.5 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.

Storage

This material is provided as lyophilized solid that is stable for 1 year at -20°C, desiccated and protected from light. Phalloidin conjugates should be reconstituted 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, 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.

Product Description

DyLight™ 488 Phalloidin allows researchers to fluorescently label the cytoskeleton of fixed cells through the binding of phalloidin to F-actin. This product is not intended for use on live cells due to the toxicity associated with phalloidin. After reconstitution the stock solution provides enough material to perform 600 assays based on a 1:40 dilution and a 100 μl assay volume.

DyLight™ 488 Fluorescent Properties: Excitation: 495 nm, Emission: 521 nm.

Background

Actin is a ubiquitous eukaryotic protein and a 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 to control cell structure and motility (1). Actin exists mainly as a fibrous polymer called 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 G-actin form (2). Phalloidin is a naturally occurring toxic bicyclic peptide found in the deathcap toadstool, Amanita phalloides, which 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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
DRAQ5 is a registered trademark of Biostatus Limited.
DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.
ProLong is a registered trademark of Life Technologies Corporation.
To Purchase # 12935S
Product # Size Price
12935S
300 units $ 388