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A3853 Sigma-Aldrich

Anti-Actin antibody , Mouse monoclonal

clone AC-40, purified from hybridoma cell culture

  •  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)   immunocytochemistry: 10-20 μg/mL using chicken or human fibroblasts by methanol/acetone fixation
  immunohistochemistry: suitable
  indirect ELISA: suitable
  microarray: suitable
  western blot: 0.5 μg/mL using chicken or human fibroblasts
form   buffered aqueous solution
shipped in   dry ice
storage temp.   −20°C
packaging   antibody small pack of 25 μL
antibody form   purified from hybridoma cell culture
Quality Level   200
antibody product type   primary antibodies

Description

General description

Actin is a highly conserved protein that is a major component of both the cytoskeletal and contractile structures in all cell types. It varies in amount, being related to the type of differentiation and to the functional state of cells and tissues. Actin can be found in two different forms of aggregation, the globular or the fibrillar state, and at least six distinct isoforms occur in vertebrates. The actins exhibit over 90% sequence homology, but each isoform has a unique NH2-terminal sequence. The isoforms are comprised of three α actins (skeletal, cardiac, smooth), one β actin (β-non-muscle) and two γ actins (γ smooth muscle and γ non-muscle). 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.

Specificity

Cross-reaction of monoclonal Anti-Actin antibody 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.

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.

Mouse monoclonal anti-actin antibody can be used for western blot assays. Furthermore, the product can also be used for immunocytochemistry (10-20μg/mL, using chicken or human fibroblasts by methanol/acetone fixation), microarray, immunohistochemistry and indirect ELISA.
Monoclonal Anti-Actin antibody produced in mouse has been used in the detection of actin in human bronchial epithelial cells using immunohistochemical assays. It has also been used in immunofluorescence detection in cardiomyocytes.

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

Physical form

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

Storage and Stability

For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots. Repeated freezing and thawing is not recommended. 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.

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.

Packaging

200 μL in glass insert

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. 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 2
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

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

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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, 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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