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  • : FAK100
  • : 100 assays
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Figure 1. Confocal fluorescence microscopy of focal adhesion and actin cytoskeleton in NIH-3T3 cells. (A) F-actin was detected using TRITC-conjugated...

Figure 2. Confocal fluorescence microscopy of focal adhesion and actin cytoskeleton in COS-7 cells revealed with triple labeling using TRITC-conjugat...

Actin Cytoskeleton / Focal Adhesion Staining Kit

Description:
Actin Cytoskeleton / Focal Adhesion Staining Kit
Trade Name:
Chemicon (Millipore)
Qty/Pk:
100 assays
Product Overview:
The Actin Cytoskeleton and Focal Adhesion Staining Kit consists of three components (TRITC-conjugated phalloidin, anti-Vinculin and DAPI) for the immunofluorescent staining of actin filaments in the cytoskeleton, focal contacts as well as the nucleus of the cells.

Reagents and materials supplied in the Actin Cytoskeleton and Focal Adhesion Staining Kit are sufficient for 100 tests (including controls).

Introduction

The actin cytoskeleton is a highly dynamic network composed of actin polymers and a large variety of associated proteins. The functions of the actin cytoskeleton is to mediate a variety of essential biological functions in all eukaryotic cells, including intra- and extra-cellular movement and structural support. To perform these functions, the organization of the actin cytoskeleton must be tightly regulated both temporally and spatially. Many proteins associated with the actin cytoskeleton are thus likely targets of signaling pathways controlling actin assembly. Actin cytoskeleton assembly is regulated at multiple levels, including the organization of actin monomers (G-protein) into actin polymers and the superorganization of actin polymers into a filamentous network (F-actin - the major constituent of microfilaments) [1]. This superorganization of actin polymers into a filamentous network is mediated by actin side-binding or cross-linking proteins [2, 3, 4]. Actin cytoskeleton is a dynamic structure that rapidly changes shape and organization in reponse to stimuli and cell cycle progression. Therefore, a disruption of its normal regulation may lead to cell transformations, hence cancer. Transformed cells have been shown to contain less F-actin than untransformed cells and exhibit atypical coordination of F-actin levels to the cell cycle [5]. Orientational distribution of actin filaments within a cell is an important determinant of cellular shape and motility. Therefore,

Focal adhesion and adherens junctions are membrane-associated complexes that serve as nucleation sites for actin filaments and as cross-linkers between the cell exterior, plasma membrane and actin cytoskeleton [6]. The function of focal adhesions are structural, linking the ECM on the outside to the actin cytoskeleton on the inside. They are also sites of signal transduction, initiating of signaling pathways in response to adhesion. Focal adhesions consist of integrin-type receptors that are attached to the extracellular matrix and are intracellularly associated with protein complexes containing vinculin (universal focal adhesion marker), talin, a-actinin, paxillin, tensin, zyxin and focal adhesion kinase (FAK) [7,8].

CHEMICON's Actin Cytoskeleton and Focal Adhesion Staining Kit (Catalog number FAK100) is a very sensitive immunocytochemical tool that contains fluorescent-labeled phalloidin (TRITC-conjugated phalloidin) to map the local orientation of actin filaments within cell and a mouse monoclonal antibody to Vinculin that is very specific for the staining of focal contacts in cells.
Applications:
The Actin Cytoskeleton & Focal Adhesion Staining Kit consists of TRITC-conjugated phalloidin, anti-Vinculin & DAPI for the immunofluorescent staining of actin filaments in the cytoskeleton as well as the nucleus of the cells.
Key Applications:
Immunocytochemistry
Species Reactivity:
  • Bovine
  • Human
  • Pig
  • Rabbit
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Components:
  • Vinculin Monoclonal antibody, purified clone 7F9 (Part No. 90227). One vial containing 100 μL at 1 mg/mL.
  • TRITC-conjugated Phalloidin (Part No. 90228). One vial containing 15 μg.
  • DAPI (Part No. 90229). One vial containing 100 μL.
Brand Family:
Chemicon
Host:
Mouse
Storage Conditions:
When stored at 2-8º C, the kit components are stable up to the expiration date. Do not freeze or expose to elevated temperatures. Discard any remaining reagents after the expiration date.
Product Name:
Actin Cytoskeleton / Focal Adhesion Staining Kit
Materials Required but Not Delivered:
1. Fixative, e.g. 4% paraformaldehyde

2. Permeabilizing reagent, e.g. 0.1% Triton X-100

3. Blocking solution (1% BSA in 1xPBS)

4. Fluorescent-labeled Secondary antibodies (For example CHEMICON Cat. No. AP124F)

5. 1 x Wash Buffer (e.g. 1xPBS containing 0.05% Tween-20)

6. Antifade mounting solution (For example: CHEMICON Cat. No. 5013)

7. Glass slides and coverslips

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