Anti-beta-Actin antibody, Mouse monoclonal (C15-1314-920)

General description

In staining of chicken gizzard ultrathin tissue cryosections, the antibody labels the dense bodies and longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane-associated dense plaque. It does not stain adult cardiac and skeletal muscles except for traces due to contaminations of the sample with non-muscle cells, or if embryonic tissue is being used.

The ACTB (β-actin) gene is mapped to human chromosome 7p22.1. β-Actin is the most abundant protein localized to the cytoplasm. ACTB is expressed ubiquitously. Actin is one of the most conserved eukaryotic proteins, it is expressed in mammals and birds. Four of the actin isoforms represent the differentiation markers of muscle tissues and two are found in almost all cells. Anti-β-Actin antibody, Mouse Monoclonal (mouse IgG1 isotype) is derived from the AC-15 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB/c mice immunized with a slightly modified synthetic b-cytoplasmic actin N-terminal peptide Ac-Asp-Asp-Asp-Ile-Ala-Ala-Leu-Val-Ile-Asp-Asn-Gly-Ser-Gly-Lys, conjugated to KLH.
In staining of chicken gizzard ultrathin tissue cryosections, the antibody labels the dense bodies and longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane-associated dense plaque. It does not stain adult cardiac and skeletal muscles except for traces due to contaminations of the sample with non-muscle cells, or if embryonic tissue is being used.

Specificity

Anti-β-Actin antibody, Mouse Monoclonal 1, 2 recognizes an epitope located on the N-terminal end of the β-isoform of actin.

Immunogen

slightly modified β-cytoplasmic actin N-terminal peptide, Ac-Asp-Asp-Asp-Ile-Ala-Ala-Leu-Val-Ile-Asp-Asn-Gly-Ser-Gly-Lys, conjugated to KLH.

Application

Monoclonal anti-beta-actin antibody can be used for microarray, indirect immunofluorescence, and immunohistochemical analyses. Furthermore, the product has been used for immunocytochemistry at 10-40 μg/mL using human foreskin fibroblasts. The antibody has also been used for western blot at 0.5-1 μg/mL using cell extract of human foreskin fibroblasts or chicken fibroblasts.
The antibody can be used for staining of acetone-fixed frozen sections, EM preparations, and microinjection experiments. B5, ethanol, methacam, or Bouin′s solutions can be used as fixatives. The epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding.

Monoclonal mouse anti-actin antibody was used as a loading control for western blot analysis of immunoprecipitated proteins from rat dorsal root ganglion cocultures.

Monoclonal mouse anti-actin was used as a loading control for western blot analysis of rat liver protein lysates.

The actin in cells of various species and tissue origin is very similar in their immunological and physical properties. As a consequence, it has been difficult to produce potent antisera to this protein. Therefore the availability of monoclonal antibodies to β-actin provides a specific and useful tool in studying the intracellular distribution of β-actin and the static and dynamic aspects of the cytoskeleton.
Monoclonal anti-beta-actin antibody can be used for microarray, indirect immunofluorescence, and immunohistochemical analyses. Furthermore, the product has been used for immunocytochemistry at 10-40 μg/mL using human foreskin fibroblasts. The antibody has also been used for western blot at 0.5-1 μg/mL using cell extract of human foreskin fibroblasts or chicken fibroblasts.
The antibody can be used for staining of acetone-fixed frozen sections, EM preparations, and microinjection experiments. B5, ethanol, methacam, or Bouin′s solutions can be used as fixatives. The epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding.
It has been used in Immunoblot analysis.

Biochem/physiol Actions

The two major cytoskeletal proteins implicated in cell motility are actin and myosin. Actin and myosin are constituents of many cell types and are involved in myriad of cellular process including locomotion, secretion, cytoplasmic streaming, phagocytosis, and cytokinesis.
ACTB (β-actin) is crucial for organogenesis, especially for the development of brain, kidney and heart. It is responsible for maintaining cell proliferation, migration and shape. Actin protein is essential for the formation of mature platelets. β−Actin plays a key role in embryonic development. Upregulation of β-actin stimulates membrane protrusions and helps in cellular motility. The encoded protein β-Actin mediates fibroblast migration in mouse and thereby control connective tissue tension.
Actin is a cytoskeletal protein that regulates cell motility, secretion, phagocytosis and cytokinesis. The NH2-terminal of actin may function as an antigen. This terminal may also modulate actin interactions and may associate with proteins such as myosin.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide.

Preparation Note

Storage and Stability: For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots at -20 °C. Repeated freezing and thawing, or storage in “frost-free” freezers, is not recommended. If slight turbidity occurs upon prolonged storage, clarify the solution by centrifugation before use. Working dilution samples should be discarded if not used within 12 hours.

Other Notes

To view an Actin antibody selection guide, please visit www.sigmaaldrich.com/actin.

Disclaimer

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.

biological source: mouse. Quality Level: 300. conjugate: unconjugated. antibody form: purified immunoglobulin. antibody product type: primary antibodies. clone: AC-15, monoclonal. form: buffered aqueous solution. mol wt: antigen 42 . kDa. species reactivity: sheep, carp, feline, chicken, rat, mouse, Hirudo medicinalis, rabbit, canine, pig, human, bovine, guinea pig. should not react with : Dictyostelium discoideum. technique(s): immunocytochemistry: 10-40 . μ. g/mL using human foreskin fibroblasts, immunohistochemistry (frozen sections): suitable, indirect immunofluorescence: suitable, microarray: suitable, western blot: 0.5-1 . μ. g/mL using cell extract of human foreskin fibroblasts or chicken fibroblasts.. isotype: IgG1. UniProt accession no.: P60709. application(s): research pathology. shipped in: dry ice. storage temp.: −. 20°C. target post-translational modification: unmodified. Gene Information: human ... ACTB(60)
mouse ... Actb(11461)
rat ... Actb(81822). Storage Class Code: 10 - Combustible liquids. WGK: WGK 3. Flash Point(F): Not applicable. Flash Point(C): Not applicable.