Cytoskeletal Structure Antibodies and Kits
Millipore provides antibodies and kits for studying components of the cytoskeleton, including actin, vimentin, keratin, neurofilaments, lamin and tubulin, and also provides innovative solutions to investigate signaling proteins that associate with the cytoskeleton.

The cytoskeleton consists of actin, microtubules, and intermediate filaments. Actin polymers, which mediate cell movement and intracellular dynamics, are regulated by signaling proteins, including those of the N-WASP family. Microtubules, directing movement of subcellular components, are stabilized by Rho GTPases, microtubule-associated proteins, and organization centers. IFs are a diverse family of structural proteins with equally diverse stabilizing mechanisms. Molecular motor proteins move along actin filaments or microtubules and include myosins, kinesins, and dyneins. These proteins mediate the sliding of filaments and transport organelles along cytoskeletal tracks.

For further biochemical analysis of cellular targets/receptors that associate with the cytoskeleton, browse our selection of antibodies, proteins and inhibitors.

PRODUCT HIGHLIGHT
Anti-Actin, clone C4

Illuminate the actin cytoskeleton with this actin antibody, validated for IHC, Western blot, ELISA and more. Confocal fluorescent analysis of Hela and NIH 3T3 using anti-actin clone C4 (red) shows that antibody staining colocalizes with Alexa Fluor 488 -Phalloidin (Green). Nucleus is stained with DAPI (Blue).
Actin is an abundant cytoskeletal protein whose peptide chain assumes two physical forms: globular actin and fibrous actin. In non-muscle cells, actin appears to be involved in a variety of functions, such as cell motility, protein & organelle trafficking, exocytosis, and phagocytosis.

MAB1501 & MAB1501R are pan-actin antibodies that bind to an epitope in a highly conserved region of actin; therefore, this antibody reacts with all six isoforms of vertabrate actin, including Dictyostelium discoideum and Physarum polycephalum actins (Lessard, 1988). This antibody reacts with both globular (G) and fillimentous (F) forms of actin and does not interfere with actin polymerization to form filaments, at a ratio as high as one antibody per two actin monomers. However, this antibody does increase the extent of polymerization when used at a lower ratio of antibody to actin. In addition to labeling myotubes, anti-actin stains myoblasts and fibroblasts (Lessard, 1983).


PRODUCT HIGHLIGHT
ProteoExtract® Cytoskeleton Enrichment and Isolation Kit

Western Blot of compartmental proteins extracted from HeLa cells using ProteoExtract Cytoskeleton Enrichment and Isolation Kit (cat. # 17-10195). Compartmental proteins were subjected to SDS-PAGE and proteins were transferred to a PVDF membrane. The membrane was probed with primary anti-GAPDH and anti-vimentin, followed by secondary antibodies. The blot was developed by enhanced chemiluminescence. Immunobloting results indicate that GAPDH (Panel A) is present in the cytoplasmic and nuclear fractions (Tristan C et al. (2011) Cell Signal 23: 317-323.), and the intermediate filament protein Vimentin (Panel B) is present exclusively in the cytoskeletal compartment.
Biochemical analysis of proteins that associate with and regulate the actin cytoskeleton has traditionally been difficult because many actin regulatory proteins and phosphoproteins move from the soluble cytoplasmic compartment to the insoluble actin cytoskeleton upon activation. The insolubility of these important proteins has made it difficult to study the biochemical changes, such as phosphorylation and nitrosylation, that occur upon binding to actin.

EMD Millipore’s ProteoExtract® Cytoskeleton Enrichment and Isolation kit enables gentle and selective separation of soluble cytosolic proteins from adherent cells, enabling more sensitive biochemical analysis (including Western blot and mass spectrometry) of the insoluble cytoskeleton and associated proteins in their native associations and conformations. These kits can greatly increase the ability to detect and study low abundance actin-associated proteins, which are typically masked in conventional Western blot analysis of whole cell lysates.

Advantages:
  • Fast and simple enrichment of cytoskeleton-associated proteins in 20 minutes
  • Maintains actin cytoskeleton and associated proteins in their native, adhered conformations
  • Significantly reduces background signal, while allowing for improved detection of low-abundance cytoskeleton-associated proteins

For visual analysis, see our ProteoExtract® Cytoskeleton Enrichment and Staining Kit (Catalogue No. 17-10210).