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A2066

Sigma-Aldrich

Anti-Actin antibody produced in rabbit

enhanced validation

affinity isolated antibody, buffered aqueous solution

Synonym(s):

Rabbit Anti-Actin

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About This Item

MDL number:
UNSPSC Code:
12352203
NACRES:
NA.41

biological source

rabbit

Quality Level

conjugate

unconjugated

antibody form

affinity isolated antibody

antibody product type

primary antibodies

clone

polyclonal

form

buffered aqueous solution

mol wt

antigen 42 kDa

species reactivity

wide range, vertebrates, human, slime mold, amoeba, chicken

enhanced validation

independent
Learn more about Antibody Enhanced Validation

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

UniProt accession no.

application(s)

research pathology

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

Gene Information

human ... ACTA1(58)
mouse ... Acta1(11459)
rat ... Acta1(29437)

General description

Actin, a highly conserved protein, 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 ACTA1 (actin, α 1) gene is mapped to human chromosome 1q42.13. The gene codes for actin 1, a thin filament component of skeletal muscle.

In immunoblotting, the product localizes actin in many species ranging from human skeletal muscle to amoeba. The product recognizes the 42 kDa actin band using immunoblotting techniques in human or animal tissue extracts.

Specificity

Anti-Actin specifically stains typical stress fibers in cultured chicken fibroblasts.

Immunogen

The C-terminal actin fragment attached to Multiple Antigen Peptide (MAP) backbone.

Application

May be used to study actin structure and function and to probe binding sites of actin-binding proteins.
Rabbit anti-actin antibody can be used for western blot assays. The antibody can also be used for immunohistochemistry (1:25, using human or animal tissue sections) and indirect immunofluorescence (1:40, using chicken fibroblasts) applications.
Used to study actin structure and function and to probe binding sites of actin-binding proteins. Protein lysates generated from embryonic mouse limb tissue was analyzed by western blot using actin as the loading control. Actin was detected using rabbit anti-actin at 1:8000. Actin detected by a rabbit anti-actin antibody was used as a loading control for MCF7 cell lysates. Actin was detected at 42 kDa.
Used to study actin structure and function and to probe binding sites of actin-binding proteins. Protein lysates generated from embryonic mouse limb tissue was analyzed by western blot using actin as the loading control. Actin was detected using rabbit anti-actin at 1:8000. Actin detected by a rabbit anti-actin antibody was used as a loading control for MCF7 cell lysates . Actin was detected at 42 kDa.

Biochem/physiol Actions

Actin is a cytoskeletal protein that regulates cell motility, secretion, phagocytosis and cytokinesis. The NH2-terminal of actin may function as an antigen. This terminal may also modulate actin interactions and may associate with proteins such as myosin. Actin protein is essential for the formation of mature platelets.
Mutations in ACTA1 (actin, α 1) is known to cause thin filament myopathy such as nemaline myopathy, congenital myopathy, actin myopathy, intranuclear rod myopathy, cap myopathy and core myopathy. Upregulation of the gene is observed in orthotopic liver transplantation without any earlier evidence of autoimmune liver disease. The encoded protein actin forms a network of filaments and is responsible for the shape of the cell. It provides cellular strength and promotes active movement of the cell.

Physical form

Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 1% bovine serum albumin and 15 mM sodium azide as preservative.

Storage and Stability

For continuous use, store at 2-8 °C for up to 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.

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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WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

常规特殊物品

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. What is the molecular weight of actin, Products  A2066 and A2547?

    Actin has a molecular weight of 42 kDa.

  3. Which type of actin does Product A2066, Anti-Actin antibody produced in rabbit, recognize?

    This antibody is a polyclonal antibody.  The immunogen was a C-terminal actin fragment (C11 peptide, Ser-Gly-Pro-Ser-Ile-Val-His-Arg-Lys-Cys-Phe) that is found in all actin sequences.

  4. Does Product A2066, Anti-Actin antibody produced in rabbit, react with my species?

    The sequence use to develop this antibody is conserved across many species from amoeba to vertebrates.  It should react with most samples.

  5. Are there any special considerations for immunohistochemistry of formalin-fixed, paraffin-embedded tissue sections?

    For FFPE sections, we recommend using enzymatic unmasking.

  6. Why do I have 3 bands on my western blot when using Product A2066, Anti-Actin antibody produced in rabbit?

    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).

  7. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  8. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

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

    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. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Cytoskeletal regulation of platelet formation: Coordination of F-actin and microtubules
Poulter N S and Thomas S G
The International Journal of Biochemistry & Cell Biology, 66 (2015)
The zebrafish runzel muscular dystrophy is linked to the titin gene.
Steffen L S, et al.
Developmental Biology, 309(2) (2007)
Leta S Steffen et al.
Developmental biology, 309(2), 180-192 (2007-08-07)
Titin (also called connectin) acts as a scaffold for signaling proteins in muscle and is responsible for establishing and maintaining the structure and elasticity of sarcomeres in striated muscle. Several human muscular dystrophies and cardiomyopathies have previously been linked to
Effects of doxorubicin cancer therapy on autophagy and the ubiquitin-proteasome system in long-term cultured adult rat cardiomyocytes.
Dimitrakis P, et al.
Cell and Tissue Research, 350(2), 361-372 (2012)
Severe congenital actin related myopathy with myofibrillar myopathy features.
Selcen D.
Neuromuscular Disorders, 25(6), 488-492 (2015)

Articles

Loading controls in western blotting application.

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