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A1978 Sigma-Aldrich

Anti-β-Actin antibody, Mouse monoclonal

clone AC-15, purified from hybridoma cell culture

Synonym: Monoclonal Anti-β-Actin

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Properties

Related Categories AC-AC, Actin Antibodies, Alphabetical Index, Antibodies, Antibodies for Cell Biology,
conjugate   unconjugated
clone   AC-15, monoclonal
biological source   mouse
application(s)   immunocytochemistry: 10-40 μg/mL using human foreskin fibroblasts
  immunohistochemistry (frozen sections): suitable
  indirect immunofluorescence: suitable
  microarray: suitable
  western blot: 0.5-1 μg/mL using cell extract of human foreskin fibroblasts or chicken fibroblasts.
species reactivity   guinea pig, canine, Hirudo medicinalis, feline, pig, carp, mouse, chicken, rabbit, sheep, rat, human, bovine
should not react with   Dictyostelium discoideum
mol wt   antigen mol wt 42 kDa
form   buffered aqueous solution
shipped in   dry ice
storage temp.   −20°C
antibody form   purified immunoglobulin
isotype   IgG1
antibody product type   primary antibodies
Featured Industry   Research Pathology
UniProt accession no.   P60709
Gene Information   human ... ACTB(60)
mouse ... Actb(11461)
rat ... Actb(81822)

Description

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.

Physical form

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

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.

Other Notes

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

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.

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.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
3

Documents

Certificate of Analysis

Certificate of Origin

Protocols & Articles

Articles

Antibodies to Actin Gene Family Isoforms

The Actin Gene Family Actin and myosin are the two major cytoskeletal proteins implicated in cell motility. As constituents of many cell types, actin and myosin are involved in a myriad of cellular p...
Keywords: Western blot

Antibodies to Actin and GAPDH

Actin and myosin are the two major cytoskeletal proteins implicated in cell motility. As constituents of many cell types, actin and myosin are involved in a myriad of cellular process including locom...
Leigh Gaskill
BioFiles 2008, 3.6, 10.
Keywords: Biofiles, Immunofluorescence, PAGE, Western blot

Antibody Basics

Immunoglobulins (Igs) are produced by B lymphocytes and secreted into plasma. The Ig molecule in monomeric form is a glycoprotein with a molecular weight of approximately 150 kDa that is shaped more ...
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Protocols

Western Blot Protocol | Immunoblotting Protocol

Western Blotting refers to the electrophoretic transfer of proteins from sodium dodecyl sulfate polyacrylamide gels to sheets of PVDF or nitrocellullose membrane, followed by immunodetection of prote...
Keywords: AGE, Buffers, Cell disruption, Detergents, Dialysis, Electroblotting, Electrophoresis, Enzyme activity, Gel electrophoresis, Immunoprecipitation, PAGE, Protein extraction, Purification, Sample preparations, Western blot

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Peer-Reviewed Papers
15

References

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