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A4700 Sigma-Aldrich

Monoclonal Anti-Actin antibody produced in mouse

clone AC-40, ascites fluid

Synonym: Monoclonal Anti-Actin

  •  NACRES NA.41

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Properties

Related Categories AC-AC, Actin Antibodies, Alphabetical Index, Antibodies, Antibodies for Cell Biology,
conjugate   unconjugated
clone   AC-40, monoclonal
biological source   mouse
application(s)   immunohistochemistry (frozen sections): suitable using methanol-fixed, frozen sections
  indirect ELISA: suitable
  indirect immunofluorescence: 1:200 using cultured human or chicken fibroblasts
  western blot: 1:500 using cultured human or chicken fibroblast extract
species reactivity   human, carp, Xenopus, canine, sheep, pig, mouse, rabbit, rat, hamster, chicken, snail, bovine, viper, guinea pig, goat
mol wt   antigen 42 kDa
shipped in   dry ice
storage temp.   −20°C
antibody form   ascites fluid
isotype   IgG2a
Quality Level   200
antibody product type   primary antibodies
contains   15 mM sodium azide

Description

General description

Actin and myosin are major cytoskeletal proteins. Actin is expressed majorly in six isoforms. The human γ-actin gene is mapped to human chromosome 17.
The antibody recognizes an epitope located on the C-terminal end of actin conserved in all actin isoforms. It specifically labels actin in a wide variety of tissues and species. The epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding. Zinc-formalin, B5, ethanol, methacarn, Brunnel′s or Bouin′s solutions may also be used as fixatives.

Monoclonal Anti-Actin (mouse IgG2a isotype) is derived from the AC-40 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from an immunized mouse. A synthetic actin C-terminal peptide attached to Multiple Antigen Peptide (MAP) backbone was used as the immunogen. The isotype is determined using Mouse Monoclonal Antibody Isotyping Reagents, Catalog No. ISO2.

The antibody recognizes an epitope located on the C-terminal end of actin conserved in all actin isoforms. It specifically labels actin in a wide variety of tissues and species. The epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding. Zinc-formalin, B5, ethanol, methacarn, Brunnel′s or Bouin′s solutions may also be used as fixatives.

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 cellular process including locomotion, secretion, cytoplasmic streaming, phagocytosis, and cytokinesis. Although actin is one of the most conserved eukaryotic proteins, it is expressed in mammals and birds as at least six isoforms characterized by electrophoresis and amino acid sequence analysis.

Specificity

Monoclonal Anti-Actin recognizes an epitope located on the C-terminal end of actin, but not on the N-terminal end. This epitope is conserved in all actin isoforms. The antibody specifically labels actin in a wide variety of tissues and species in immunoblotting (band at 42 kDa), in immunofluorescent staining of cultured cell lines and in immunohistology. Cross-reaction has been observed with human, bovine, sheep, goat, pig, rabbit, dog, mouse, rat, guinea pig, hamster, chicken, carp, viper, frog, and snail. The antibody may be used in staining of methanol-fixed, frozen sections, and the epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding. Zinc-formalin, B5, ethanol, methacarn, Brunnel′s or Bouin′s solution may also be used as fixatives.

Immunogen

synthetic actin C-terminal peptide, attached to a Multiple Antigen Peptide (MAP) backbone

Application

Monoclonal anti-actin antibody was used for western blot analysis of Cos-7 cell lysates to ensure equal protein loading.

Monoclonal rabbit anti-actin has been used:
• in the detection of actin in HeLa cells, breast cancer cell line and leukemia cell line using western blotting
• in human fibroblasts by immunofluorescence detection

Western blot analysis of MDCK cell lysates were performed using monoclonal anti-actin antibody as a primary antibody.

Physical form

Supplied as ascites fluid with 15mM sodium azide as a preservative.

Storage and Stability

For continuous use, store at 2-8 °C for a maximum of one month. For extended storage, the solution may be frozen in working aliquots. 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.

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.

Biochem/physiol Actions

Actin and myosin are constituents of many cell types and are involved in cellular process including locomotion, secretion, cytoplasmic streaming, phagocytosis, and cytokinesis. The N-terminal region of actin appears to be a major antigenic region, and may be involved in the interaction of actin with other proteins such as myosin. Among the six isoforms of actin, four of them represent the differentiation markers of muscle tissues and two are found practically in all cells. There are three α-actins (skeletal, cardiac, and smooth muscle), one β−actin (β-nonmuscle), and two γ-actins (γ-smooth muscle and γ-nonmuscle). Actin isoforms show >90% overall sequence homology, but only 50–60% homology in their NH2-terminal residues.
The actin in cells of various species and tissues are very similar in their immunological and physical properties characterized by electrophoresis and amino acid sequence analysis.(from bulletin) They mediate cytoskeletal remodeling during T cell activation. Mutation in the γ-actin and β-actin gene is associated with a developmental disorder called Baraitser-Winter syndrome. Mutation in the γ-actin leads to hearing loss.

Other Notes

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

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 3
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable
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

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 ...
Keywords: Affinity chromatography, Centrifugation, Chromatography, Digestions, Direct immunofluorescence, Gene expression, High performance liquid chromatography, Immunofluorescence, Ion Exchange, Microscopy, Precipitation, Purification, Rheumatology, Scanning electron microscopy

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, Detection methods, 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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