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Anti-Actin antibody, Mouse monoclonal

Name: Anti-Actin antibody, Mouse monoclonal
Brand: SIGMA

clone AC-40, purified from hybridoma cell culture

Synonym(s):

Anti-ACTA

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About This Item

NACRES:

NA.41

UNSPSC Code:

12352203

Conjugate:

unconjugated

Clone:

AC-40, monoclonal

Application:

ARR, ELISA (i), ICC, IHC, WB

Citations:

606

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Properties

biological source

mouse

Quality Segment

200

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

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.

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[1] 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.

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.

Physical form

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

Preparation Note

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.

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This Item	A2066	A5228	A5316
Sigma-Aldrich A3853 Anti-Actin antibody, Mouse monoclonal	Sigma-Aldrich A2066 Anti-Actin antibody produced in rabbit	Sigma-Aldrich A5228 Anti-Actin, α-Smooth Muscle antibody, Mouse monoclonal	Sigma-Aldrich A5316 Monoclonal Anti-β-Actin antibody produced in mouse
conjugate unconjugated	conjugate unconjugated	conjugate unconjugated	conjugate unconjugated
antibody form purified from hybridoma cell culture	antibody form affinity isolated antibody	antibody form purified immunoglobulin	antibody form ascites fluid
clone AC-40, monoclonal	clone polyclonal	clone 1A4, monoclonal	clone AC-74, monoclonal
biological source mouse	biological source rabbit	biological source mouse	biological source mouse
Quality Level 200	Quality Level 200	Quality Level 200	Quality Level 200
technique(s) immunocytochemistry: 10-20 μg/mL using chicken or human fibroblasts by methanol/acetone fixation, indirect ELISA: suitable, western blot: 0.5 μg/mL using chicken or human fibroblasts, immunohistochemistry: suitable, microarray: suitable	technique(s) immunohistochemistry (formalin-fixed, paraffin-embedded sections): 1:25 using human or animal tissue sections, indirect immunofluorescence: 1:40 using chicken fibroblasts, western blot: 1:100 using chicken gizzard extract	technique(s) immunocytochemistry: suitable using smooth muscle cells, immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable using smooth muscle cells, indirect ELISA: suitable, indirect immunofluorescence: 5-10 μg/mL using blood vessels in formalin-fixed, paraffin-embedded human tonsil or appendix tissue sections, microarray: suitable, western blot: suitable using smooth muscle cells	technique(s) immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable, indirect ELISA: suitable, indirect immunofluorescence: 1:1,000 using cultured human or chicken fibroblasts, western blot: 1:5,000 using cultured human or chicken fibroblast cell extracts

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Storage Class

10 - Combustible liquids

wgk

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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