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A5441

Anti-β-Actin (ACTB) Antibody

mouse monoclonal, AC-15

Monoclonal Anti-β-Actin antibody produced in mouse

Synonym(s):

Monoclonal Anti-β-Actin, Anti beta actin monoclonal antibody

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100 μL
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$588.00
0.2 mL
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$700.00
0.5 mL
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$1,350.00

About This Item

UNSPSC Code:
12352203
NACRES:
NA.41
Conjugate:
unconjugated
Clone:
AC-15, monoclonal
Application:
ELISA (i), IF, IHC (p), WB
Citations:
10789

$588.00


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Product Name

Monoclonal Anti-β-Actin antibody produced in mouse, clone AC-15, ascites fluid

biological source

mouse

Quality Level

conjugate

unconjugated

antibody form

ascites fluid

antibody product type

primary antibodies

clone

AC-15, monoclonal

mol wt

antigen 42 kDa

contains

15 mM sodium azide

species reactivity

sheep, carp, feline, chicken, rat, mouse, Hirudo medicinalis, rabbit, canine, pig, human, bovine, guinea pig

should not react with

Dictyostelium discoideum

technique(s)

immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable, indirect ELISA: suitable, indirect immunofluorescence: 1:1,000-1:2,000 using cultured human or chicken fibroblasts, western blot: 1:5,000-1:10,000 using cultured human or chicken fibroblast cell extracts

isotype

IgG1

UniProt accession no.

application(s)

research pathology

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

Gene Information

human ... ACTB(60)
mouse ... Actb(11461)
rat ... Actb(81822)

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.
Monoclonal Anti-β-Actin (mouse IgG1 isotype) is derived from the AC-15 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from an immunized mouse. Actin is one of the most conserved eukaryotic proteins, it is expressed in mammals and birds as at least six isoforms. Four of them represent the differentiation markers of muscle tissues and two are found practically in all cells. There are three α-actins (α-skeletal, α-cardiac, and α-smooth muscle), one β-actin (β-nonmuscle), and two γ-actins (γ-smooth muscle and γ-non-muscle). Actin isoforms show >90% overall sequence homology, but only 50−60% homology in their 18 NH2-terminal residues. The NH2-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. 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-fixed and paraffin-embedding.

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-β-Actin antibody produced in mouse has also been used:
  • in immunofluorescence staining
  • in immunoblotting
  • in immunohistochemistry
  • as a control for protein arrays
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.

Biochem/physiol Actions

Actin and myosin are constituents of many cell types and are involved in a myriad of cellular processes including locomotion, cytokinesis, cytoplasmic streaming, secretion and phagocytosis. The actin in cells of various species and tissue origin is very similar in its immunological and physical properties.
Monoclonal Anti β-Actin antibody recognizes an epitope located on the N-terminal end of the β-isoform of actin. The antibody specifically labels β-actin in a wide variety of tissues and species using immunoblotting (42 kDa), immunofluorescent staining of cultured cell lines, and immunohistochemistry.

Physical form

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

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
A5316A2228A1978
Gene Information

human ... ACTB(60)
mouse ... Actb(11461)
rat ... Actb(81822)

Gene Information

human ... ACTB(60)
mouse ... Actb(11461)
rat ... Actb(81822)

Gene Information

human ... ACTB(60)
mouse ... Actb(11461)
rat ... Actb(81822)

Gene Information

human ... ACTB(60)
mouse ... Actb(11461)
rat ... Actb(81822)

clone

AC-15, monoclonal

clone

AC-74, monoclonal

clone

AC-74, monoclonal

clone

AC-15, monoclonal

biological source

mouse

biological source

mouse

biological source

mouse

biological source

mouse

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

conjugate

unconjugated

species reactivity

sheep, carp, feline, chicken, rat, mouse, Hirudo medicinalis, rabbit, canine, pig, human, bovine, guinea pig

species reactivity

rabbit, sheep, cat, guinea pig, bovine, mouse, chicken, wide range, Hirudo medicinalis, canine, Drosophila, carp, pig, rat, human

species reactivity

rabbit, sheep, cat, guinea pig, bovine, mouse, chicken, wide range, Hirudo medicinalis, canine, Drosophila, carp, pig, rat, human

species reactivity

sheep, carp, feline, chicken, rat, mouse, Hirudo medicinalis, rabbit, canine, pig, human, bovine, guinea pig

antibody form

ascites fluid

antibody form

ascites fluid

antibody form

purified immunoglobulin

antibody form

purified immunoglobulin


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Storage Class

10 - Combustible liquids

wgk

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable



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Certificates of Analysis (COA)

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Protocols

Tips and troubleshooting for FFPE and frozen tissue immunohistochemistry (IHC) protocols using both brightfield analysis of chromogenic detection and fluorescent microscopy.

Western blot protocol details protein transfer from gels to nitrocellulose, crucial for immunoblotting procedures in research.

Articles

Antibodies combine with specific antigens to generate an exclusive antibody-antigen complex. Learn about the nature of this bond and its use as a molecular tag for research.

Learn differences in monoclonal vs polyclonal antibodies including how antibodies are generated, clone numbers, and antibody formats.

Immunofluorescence uses antibody-conjugated fluorescent molecules for protein localization, modification confirmation, and protein complex visualization.

View All Articles

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Global Trade Item Number

SKUGTIN
A5441-.2ML04061838443557
A5441-100UL04061838443571
A5441-.5ML04061838443564

Questions

1–8 of 8 Questions  
  1. species of antigen

    1 answer
    1. This is not a species specific antigen, it is a 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. It has multi-species reactivity as it is a highly conserved region of the protein among species.

      Helpful?

  2. What is the Department of Transportation shipping information for this product?

    1 answer
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

      Helpful?

  3. What is the antibody concentration of Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    1 answer
    1. This antibody is ascities fluid.  The amount of protein present and the amount of IgG1 present is indicated on the Lot-specific Certificate of Analysis.

      Helpful?

  4. Can I use Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse, to detect mouse beta actin?

    1 answer
    1. This antibody will detect mouse beta actin.  When used for western blotting, the secondary antibody may react with mouse IgG from the sample resulting with a bands at 50 kDa and 25 kDa (reducing conditions) or approximately 150 kDa (non-reducing conditions).  It is best to use a directly conjugated antibody for immunohistochemistry or immunofluorescent assays.  We offer Product numbers A3854, peroxidase conjugated.

      Helpful?

  5. What is the molecular weight of Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    1 answer
    1. Beta Actin has a molecular weight of 42 kDa.

      Helpful?

  6. What IgG light chain (kappa or lambda) is associated with Clone AC-15 for Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    1 answer
    1. We have not determined the light chain used by monoclonal antibody AC-15.  95% of mouse antibodies have kappa light chains.

      Helpful?

  7. Why do I have 3 bands on my western blot when using Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    1 answer
    1. While actin should run as a 42 kDa band when detected by Western blotting, there are recent observations that cleavage products can be seen in cells undergoing apoptosis. Initial cleavage gives a band at 41 kDa, which may appear as a doublet on the blot. In addition, further cleavage results in a major band at 30 kDa and a small band at 14 kDa which have been shown to be cleavage products due to the action of interleukin 1 -converting enzyme. This may explain the detection of smaller bands when blotting with this antibody. This banding pattern may be detected with other actin antibodies as well. Kayalar, C., et al., Proc. Natl. Acad. Sci., USA, 93:2234-2238, (1996).

      Helpful?

  8. What peroxidase-conjugated secondary antibody should I use with Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    1 answer
    1. We recommend using Product No. A9044, rabbit anti-mouse IgG, peroxidase conjugate as a secondary antibody. We also offer this same antibody directly conjugated to peroxidase as Product No. A3854.

      Helpful?

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