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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| A | ELISA, ICC, IF, IHC, IH(P), WB | M | Ascites | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB1501 |
| Replaces | 04-1040 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | Anti-Actin Antibody, clone C4 |
| Alternate Names |
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| Background Information | Actins are ubiquitous eukaryotic proteins that serve as multi-functional, basic building blocks of cytoskeletal microfilaments. They play critical roles in a wide range of cellular processes, including cell division, cell migration, chromatin remodeling, transcriptional regulation, and vesicle trafficking. These functions are attributed to their ability to form filaments, which can quickly assemble and disassemble depending upon the needs of the cell. At least six different actin types have been reported in mammals. Although actins show about 90% overall sequence homology, isoforms do show spatial, temporal, and tissue-specific expression patterns and only 50-60% homology is found in their 18 N-terminal residues. Beta- and gamma-actins, also known as cytoplasmic actins, are highly conserved in higher animals and are predominantly expressed in non-muscle cells where they control cell structure and motility. They are nearly identical proteins and differ only in four amino acids at the N-terminal region. The other four actin isoforms are typically found in specific adult muscle tissue types. alpha-cardiac and alpha-skeletal actins are expressed in striated cardiac and skeletal muscles, respectively. Alpha and gamma actins are primarily found in vascular smooth muscle and enteric smooth muscles, respectively. It has been shown that under calcium-bound conditions, beta-actin exhibits more dynamic behavior than gamma-actin with higher rates of polymerization and depolymerization. Also, beta- and gamma-actins can readily copolymerize, and the resulting filaments exhibit polymerization and depolymerization rates that vary depending on the ratio of beta- to gamma-actin (Lessard, JL., et al (1988). Cell Motility Cytoskeleton 10(3); 349-362). |
| Product Information | |
|---|---|
| Format | Ascites |
| Control |
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| Presentation | Mouse monoclonal Ascites fluid, with 0.01% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Reliably and specifically detect actin using this Anti-Actin Antibody, clone C4. This highly published monoclonal antibody is validated for use in ELISA, IC, IF, IH, IH(P) & WB. This mAb is also available as a fluorescent conjugate. |
| Key Applications |
|
| Application Notes | Indirect immunofluorescence at 1:100: Tissue culture cells -- fix with formaldehyde, treat with methanol or acetone. Glycerinated myofibrils -- fix fibers with formaldehyde, treat with cold methanol. Stains I-bands intensely and stress fibers in human fibroblasts. Cryostat sections (6 µm) -- quick frozen in isopentane, slides treated with gelatin and formaldehyde. Immunoblots: 1:100-1:1,000 (Otey, 1987):On muscle homogenates subject to SDS-PAGE, reacts relatively uniformly with a 43 kD protein present in skeletal, cardiac, gizzard and aorta tissues. Appears to react with all isoforms of actin found in these preparations and shows a strong reaction with the alpha-actin found in skeletal, cardiac, and arterial muscle. Iodination (Lessard, 1979). Solid phase binding assay ELISA: 1:800-1:1,000 dilution from a previous lot was shown to be strongly reactive with cytoplasmic actin and shows a significant binding to gizzard, skeletal, arterial and cardiac actins. Also shows a significant binding to both Dictyostelium discoidum and Physarum polycephalum. ELISA: strongly reactive with the cytoplasmic actin and shows a significant binding to gizzard, skeletal, arterial and cardiac actins. Also shows a significant binding to both Dictyostelium discoidum and Physarum polycephalum. Optimal working dilutions must be determined by end user. |
| Biological Information | |
|---|---|
| Immunogen | Purified chicken gizzard actin (Lessard, 1988). |
| Epitope | Epitope is conserved in all known actins. |
| Clone | C4 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | MAB1501 is a pan-actin antibody that binds to an epitope in a highly conserved region of actin; therefore, this antibody reacts with all six isoforms of vertabrate actin (Lessard, 1988). 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. Although clone C4 is prepared as an antibody to chicken gizzard muscles actin, it reacts with actins from all vertebrates, as well as with Dictyostelium discoideum and Physarum polycephalum actins (Lessard, 1988). |
| Isotype | IgG1κ |
| Species Reactivity |
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| Species Reactivity Note | To date, all animal species and cell types with an actin form react by indirect immunofluorescence or immunoblot, including plant actin. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The product encoded by this gene belongs to the actin family of proteins, which are highly conserved proteins that play a role in cell motility, structure and integrity. Alpha, beta and gamma actin isoforms have been identified, with alpha actins being a major constituent of the contractile apparatus, while beta and gamma actins are involved in the regulation of cell motility. This actin is an alpha actin that is found in skeletal muscle. Mutations in this gene cause nemaline myopathy type 3, congenital myopathy with excess of thin myofilaments, congenital myopathy with cores, and congenital myopathy with fiber-type disproportion, diseases that lead to muscle fiber defects. |
| Gene Symbol |
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| Purification Method | Unpurified |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P68133 # Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells. SIZE: 377 amino acids; 42051 Da SUBUNIT: Polymerization of globular actin (G-actin) leads to a structural filament (F-actin) in the form of a two-stranded helix. Each actin can bind to 4 others. Interacts with TTID. SUBCELLULAR LOCATION: Cytoplasm, cytoskeleton. DISEASE: SwissProt: P68133 # Defects in ACTA1 are the cause of nemaline myopathy type 3 (NEM3) [MIM:161800]. Nemaline myopathy (NEM) is a form of congenital myopathy characterized by abnormal thread- or rod-like structures in muscle fibers on histologic examination. The clinical phenotype is highly variable, with differing age at onset and severity. & Defects in ACTA1 are a cause of congenital myopathy with excess of thin myofilaments (CM) [MIM:102610]. & Defects in ACTA1 are a cause of congenital myopathy with fiber-type disproportion (CFTD) [MIM:255310]; also known as congenital fiber-type disproportion myopathy (CFTDM). CFTD is a genetically heterogeneous disorder in which there is relative hypotrophy of type 1 muscle fibers compared to type 2 fibers on skeletal muscle biopsy. However, these findings are not specific and can be found in many different myopathic and neuropathic conditions. SIMILARITY: SwissProt: P68133 ## Belongs to the actin family. MISCELLANEOUS: In vertebrates 3 main groups of actin isoforms, alpha, beta and gamma have been identified. The alpha actins are found in muscle tissues and are a major constituent of the contractile apparatus. The beta and gamma actins coexist in most cell types as components of the cytoskeleton and as mediators of internal cell motility. |
| Molecular Weight | 43 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Routinely evaluated by Western Blot on A431 lysates. Western Blot Analysis: 1:500 dilution of this lot detected ACTIN on 10 ug of A431 lysates. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable at -20°C in undiluted aliquots for up to 12 months from date of receipt. Do not store in a diluted format. Avoid repeated freeze/thaw cycles. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |