The cytoskeleton provides support for the cell, components of the intracellular transport, and cell division. The cytoskeleton consists of three different protein filaments: microtubules, intermediate filaments, and microfilaments (actin).
Alpha/beta tubulin heterodimers polymerize to form microtubules. Microtubules are involved in many cell functions including maintenance of cell structure, intracellular transport, and the formation of mitotic spindles during cell division.
Intermediate filaments are composed of keratin heterodimers which consist of an acidic keratin and a basic keratin. The tissue in which they are expressed determines in part the primary types of intermediate filaments. Cytokeratins are expressed in epithelial cells, vimentin in cells of mesenchyme origin, while desmin is expressed in skeletal, visceral and some vascular smooth muscle cells.
Actin makes up the majority of the cytoskeleton, with at least six isoforms known in mammals. Actin filaments are composed of double helical cylindrical tubes of alpha actin or beta actin. Alpha actin is expressed in muscle cells, while beta actin and gamma actin are expressed primarily in non-muscle cells.
Cytoskeleton marker antibodies can also help elucidate the roles proteins may play in tasks that are centered in or influenced by the cytoskeleton. Invitrogen cytoskeleton marker antibodies are designed to dependably detect the key cytoskeleton targets. Each antibody is validated for use in various applications. Key cytoskeleton marker targets include:
Immunofluorescent analysis of actin in HeLa cells. An actin polyclonal antibody (Cat. No. PA5-11570) was used at a dilution of 1:10–50, followed by a fluor-conjugated goat anti-rabbit secondary antibody (green). Nuclei were stained with DAPI (blue).
Immunofluorescence analysis of alpha Tubulin was performed using 70% confluent log phase LNCaP cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with alpha Tubulin (YL1/2) Rat Monoclonal Antibody at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rat IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 (Cat. No. A-11006) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade Gold Antifade Mountant with DAPI (Cat. No. S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Cat. No. R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
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