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A3853

Sigma-Aldrich

Anti-Actin antibody, Mouse monoclonal

clone AC-40, purified from hybridoma cell culture

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Synonym(s):
Anti-ACTA
NACRES:
NA.41

biological source

mouse

Quality Level

conjugate

unconjugated

antibody form

purified from hybridoma cell culture

antibody product type

primary antibodies

clone

AC-40, monoclonal

form

buffered aqueous solution

packaging

antibody small pack of 25 μL

technique(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

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

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This Item
A5316A2228A2066
biological source

mouse

biological source

mouse

biological source

mouse

biological source

rabbit

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

clone

AC-40, monoclonal

clone

AC-74, monoclonal

clone

AC-74, monoclonal

clone

polyclonal

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

packaging

antibody small pack of 25 μL

packaging

-

packaging

-

packaging

-

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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.

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.

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.

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.

Storage Class

10 - Combustible liquids

wgk_germany

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)


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The actin gene family: function follows isoform. The actin gene family: function follows isoform.
Perrin BJ and Ervasti JM
Cytoskeleton (Hoboken, N.J.), 67(10), 630-634 (2010)
In vivo and in vitro effects of two novel gamma-actin (ACTG1) mutations that cause DFNA20/26 hearing impairment.
Morin M, et al.
Human Molecular Genetics, 18(16), 3075-3089 (2009)
Disruption of a GATA4/Ankrd1 signaling axis in cardiomyocytes leads to sarcomere disarray: implications for anthracycline cardiomyopathy.
Chen B, et al.
PLoS ONE, 7(4), e35743-e35743 (2012)
Actin structure and function.
Dominguez R and Holmes KC
Annual Review of Biophysics, 40, 169-186 (2011)
De novo mutations in the actin genes ACTB and ACTG1 cause Baraitser-Winter syndrome.
Riviere JB, et al.
Nature Genetics, 44(4), 440-440 (2012)

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