Date published: 2025-9-5
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α-Actin Antibody (1A4) - Western Blotting - Image 396211
α-Actin Antibody (1A4) Alexa Fluor® 647: sc-32251 AF647. Direct fluorescent western blot analysis of α-Actin expression in C2C12 (A), A-10 (B), BC3H1 (C), Sol8 (D) and L6 (E) whole cell lysates. Blocked with UltraCruz® Blocking Reagent: sc-516214. Cruz Marker™ Molecular Weight Standards detected with Cruz Marker™ MW Tag-Alexa Fluor® 488: sc-516790.
α-Actin Antibody (1A4) Alexa Fluor® 647: sc-32251 AF647. Direct fluorescent western blot analysis of α-Actin expression in C2C12 (A), A-10 (B), BC3H1 (C), Sol8 (D) and L6 (E) whole cell lysates. Blocked with UltraCruz® Blocking Reagent: sc-516214. Cruz Marker™ Molecular Weight Standards detected with Cruz Marker™ MW Tag-Alexa Fluor® 488: sc-516790.

α-Actin Antibody (1A4): sc-32251

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Datasheets
  • α-Actin Antibody (1A4) is a mouse monoclonal IgG2a κ α-Actin antibody, cited in 414 publications, provided at 200 µg/ml
  • raised against an N-terminal decapeptide of α-Actin of human origin
  • alpha-Actin Antibody (1A4) is recommended for detection of smooth muscle α-Actin of mouse, rat and human origin by WB, IP, IF, IHC(P) and ELISA; non cross-reactive with actin from fibroblasts (β-and γ-cytoplasmic), myocardium (α-myocardial), and striated muscle (α-sarcomeric)
  • Anti-alpha-Actin Antibody (1A4) is available conjugated to agarose for IP; HRP for WB, IHC(P) and ELISA; and to either phycoerythrin or FITC for IF, IHC(P) and FCM
  • also available conjugated to Alexa Fluor® 488, Alexa Fluor® 546, Alexa Fluor® 594 or Alexa Fluor® 647 for WB (RGB), IF, IHC(P) and FCM, and for use with RGB fluorescent imaging systems, such as iBright™ FL1000, FluorChem™, Typhoon, Azure and other comparable systems
  • also available conjugated to Alexa Fluor® 680 or Alexa Fluor® 790 for WB (NIR), IF and FCM; for use with Near-Infrared (NIR) detection systems, such as LI-COR®Odyssey®, iBright™ FL1000, FluorChem™, Typhoon, Azure and other comparable systems
  • Contact our Technical Service Department (or your local Distributor) for more information on how to receive a FREE 10 µg sample of α-Actin (1A4): sc-32251.
  • m-IgG Fc BP-HRP, m-IgG2a BP-HRP and m-IgGκ BP-HRP are the preferred secondary detection reagents for α-Actin Antibody (1A4) for WB and IHC(P) applications. These reagents are now offered in bundles with α-Actin Antibody (1A4) (see ordering information below).

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SEE ALSO...

Additional Actin Antibodies including β-Actin, γ-Actin, Muscle Actin and Smooth Muscle Actin

α-Actin Antibody (1A4) is a mouse monoclonal IgG2a kappa light chain antibody that detects alpha-Actin protein of mouse, rat, and human origin by western blotting (WB), immunoprecipitation (IP), immunofluorescence (IF), immunohistochemistry with paraffin-embedded sections (IHCP), and enzyme-linked immunosorbent assay (ELISA). Anti-alpha-Actin antibody (1A4) is available in both non-conjugated and various conjugated forms, including agarose, horseradish peroxidase (HRP), phycoerythrin (PE), fluorescein isothiocyanate (FITC), and multiple Alexa Fluor® conjugates. Actin plays a vital role in cellular structure and function as a key component of the cytoskeleton, providing mechanical support and shape to cells. Alpha-Actin forms dynamic filaments in the cytoplasm that are essential for various cellular processes, including motility, division, and intracellular transport. Rapid polymerization and depolymerization of alpha-Actin allows cells to adapt shape and movement in response to environmental cues, making this protein crucial for processes such as wound healing and immune responses. Alpha-Actin contributes significantly to the contractile apparatus in muscle cells, enabling muscle contraction and relaxation. Members of the small GTPase family tightly control alpha-Actin dynamics by orchestrating the assembly and disassembly of actin filaments, ensuring proper cellular function and response to stimuli.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.

Alexa Fluor® is a trademark of Molecular Probes Inc., OR., USA

LI-COR® and Odyssey® are registered trademarks of LI-COR Biosciences

α-Actin Antibody (1A4) References:

  1. Muscle disease caused by mutations in the skeletal muscle alpha-actin gene (ACTA1).  |  Sparrow, JC., et al. 2003. Neuromuscul Disord. 13: 519-31. PMID: 12921789
  2. Cardiac α-actin over-expression therapy in dominant ACTA1 disease.  |  Ravenscroft, G., et al. 2013. Hum Mol Genet. 22: 3987-97. PMID: 23736297
  3. Nuclear Smooth Muscle α-actin in Vascular Smooth Muscle Cell Differentiation.  |  Kwartler, CS., et al. 2023. Res Sq.. PMID: 36909460
  4. Smooth muscle α-actin missense variant promotes atherosclerosis through modulation of intracellular cholesterol in smooth muscle cells.  |  Kaw, K., et al. 2023. Eur Heart J. 44: 2713-2726. PMID: 37377039
  5. Nuclear Smooth Muscle α-actin Participates in Vascular Smooth Muscle Cell Differentiation.  |  Kwartler, CS., et al. 2023. Nat Cardiovasc Res. 2: 937-955. PMID: 38919852
  6. Role of microtubule-associated proteins in the control of microtubule assembly.  |  Maccioni, RB. and Cambiazo, V. 1995. Physiol Rev. 75: 835-64. PMID: 7480164
  7. Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia.  |  Nobes, CD. and Hall, A. 1995. Cell. 81: 53-62. PMID: 7536630
  8. A discourse on modeling F-actin.  |  Schutt, CE., et al. 1995. J Struct Biol. 115: 186-98. PMID: 7577239
  9. Actin cytoskeleton. Setting the pace of cell movement.  |  Barkalow, K. and Hartwig, JH. 1995. Curr Biol. 5: 1000-2. PMID: 8542272
  10. The origins and evolution of eukaryotic proteins.  |  Doolittle, RF. 1995. Philos Trans R Soc Lond B Biol Sci. 349: 235-40. PMID: 8577833
  11. Elastic fibres are an essential component of human placental stem villous stroma and an integrated part of the perivascular contractile sheath.  |  Graf, R., et al. 1996. Cell Tissue Res. 283: 133-41. PMID: 8581953
  12. Actin bundles in human hair follicles as revealed by confocal laser microscopy.  |  Furumura, M. and Ishikawa, H. 1996. Cell Tissue Res. 283: 425-34. PMID: 8593672

Ordering Information

Product NameCatalog #UNITPriceQtyFAVORITES

α-Actin Antibody (1A4)

sc-32251
200 µg/ml
$316.00

α-Actin Antibody (1A4): m-IgG Fc BP-HRP Bundle

sc-528446
200 µg Ab; 10 µg BP
$354.00

α-Actin Antibody (1A4): m-IgGκ BP-HRP Bundle

sc-520826
200 µg Ab, 40 µg BP
$354.00

α-Actin Antibody (1A4): m-IgG2a BP-HRP Bundle

sc-547102
200 µg Ab; 10 µg BP
$354.00

α-Actin Antibody (1A4) AC

sc-32251 AC
500 µg/ml, 25% agarose
$416.00

α-Actin Antibody (1A4) HRP

sc-32251 HRP
200 µg/ml
$316.00

α-Actin Antibody (1A4) FITC

sc-32251 FITC
200 µg/ml
$330.00

α-Actin Antibody (1A4) PE

sc-32251 PE
200 µg/ml
$343.00

α-Actin Antibody (1A4) Alexa Fluor® 488

sc-32251 AF488
200 µg/ml
$357.00

α-Actin Antibody (1A4) Alexa Fluor® 546

sc-32251 AF546
200 µg/ml
$357.00

α-Actin Antibody (1A4) Alexa Fluor® 594

sc-32251 AF594
200 µg/ml
$357.00

α-Actin Antibody (1A4) Alexa Fluor® 647

sc-32251 AF647
200 µg/ml
$357.00

α-Actin Antibody (1A4) Alexa Fluor® 680

sc-32251 AF680
200 µg/ml
$357.00

α-Actin Antibody (1A4) Alexa Fluor® 790

sc-32251 AF790
200 µg/ml
$357.00

I wonder if this antibody can be used to identify muscle cells？

Asked by: Anonymous
Thank you for your question. alpha-Actin Antibody (1A4) is recommended for detection of smooth muscle α-Actin of mouse, rat and human origin by WB, IP, IF, IHC(P) and ELISA; non cross-reactive with actin from fibroblasts (β-and γ-cytoplasmic), myocardium (α-myocardial), and striated muscle (α-sarcomeric).
Answered by: Technical Service SH
Date published: 2023-03-17

I failed in ELISA with this antibody. Briefly, I coated alpha-actin on Ni plate, applied this antibody HRP, substrate TMB. No color.

Asked by: LucyLu
Thank you for your question. Please contact our Technical Service Department for assistance in troubleshooting this antibody.
Answered by: Technical Support
Date published: 2020-07-07

Quel est le tampon de démasquage et la concentration utilisé pour un marquage sur coupe paraffine ?

Asked by: acertain
Merci pour votre question. Sur coupes en paraffine, nous recommandons d’effectuer un démasquage de l’antigène en tampon citrate. Vous trouverez plus de précisions dans notre page protocoles dans le lien suivant : https://www.scbt.com/scbt/resources/protocols Thank you for your question. In IHC on paraffin embedded tissues sections, we recommend to perform antigen retrieval in citrate buffer. You will find additional information in our protocols in the following link : https://www.scbt.com/scbt/resources/protocols
Answered by: SCBT Heidelberg Support
Date published: 2018-11-27

If I want to perform a Western blot using the α-Actin Antibody (1A4) on human samples, what secondary reagent should I use for detection?

Asked by: Trav11
We recommend using one of our exclusive Mouse IgG Binding Proteins as a secondary detection reagent, such as m-IgGκ BP-HRP: sc-516102, for Western blot. A complete list of all available binding proteins is available on our website here: https://www.scbt.com/scbt/browse/support-products-mouse-igg-binding-proteins/_/N-ecrety
Answered by: Tech Service 8
Date published: 2016-12-16

Can I use this mouse monoclonal antibody for triple staining even if my other primary antibodies were also raised in mice?

Asked by: Trav11
When looking to perform triple staining with multiple mouse primary antibodies, we recommend using our directly conjugated mouse monoclonal primary antibodies. The α-Actin Antibody (1A4): sc-32251 is currently available conjugated to HRP (sc-32251 HRP), phycoerythrin (sc-32251 PE), fluorescein (sc-32251 FITC), Alexa Fluor® 488 (sc-32251 AF488) or Alexa Fluor® 647 (sc-32251 AF647).
Answered by: Tech Service 8
Date published: 2016-12-16

I would like to know whether this antibody is usable to evaluate the expression pattern of alfa-actin protein in adult zebrafish especially skeletal muscles. If not, could you please tell us the other useful antibodies?

Asked by: Yosuke
Thank you for your question. α-Actin Antibody (1A4): sc-32251 is recommended for detection of smooth muscle α-Actin, rather than skeletal muscle actin isoforms. This antibody has not yet been tested in zebrafish, and it is unknown if it would work for that species. As a recommended alternative, we would suggest our mouse monoclonal, β-Actin Antibody (C4): sc-47778. This antibody is recommended for detection of β-Actin; may cross-react with all six known isoforms of Actin (including cytoplasmic β- and γ- Actin isoforms, skeletal, cardiac, and vascular α-Actin isoforms, and enteric γ-Actin isoform). This antibody has also been cited in the literature used in zebrafish in the following articles on NCBI's PubMed: PMID: 25075304, 22139372.
Answered by: Technical Support
Date published: 2016-12-02
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Rated 5 out of 5 by from Great for WBIt is a good antibody in WB. It works in dilution 1:1000.
Date published: 2024-03-19
Rated 5 out of 5 by from α-SMA specific antibodyThe antibody was successfully used in IF and flow cytometry during mouse embryonic stem cell differentiation.
Date published: 2024-02-27
Rated 5 out of 5 by from alpha-SMAThis antibody was used successfully for Western blot (1:2000) with clear background.
Date published: 2022-09-26
Rated 4 out of 5 by from Good AntibodyThe antibody works perfectly for immuofluorescence on human cardiac fibroblast activated with TGFBeta
Date published: 2022-03-04
Rated 5 out of 5 by from spécifique et croise avec des espèces de renteJ'utilise cet anticorps depuis quelques années car il croise avec l'espèce bovine et donne des résultats spécifiques
Date published: 2022-01-07
Rated 5 out of 5 by from very clean backgroundWB recommends the use of 1:1000 dilution, antibody specific binding and strong band appearance. So far, no other experiments have been done.
Date published: 2017-11-01
Rated 1 out of 5 by from Not well work in WBToo many unspecific WB band detected in human liver cancer BEL-7404 lysates. dilution ratio is 1:100
Date published: 2017-10-11
Rated 4 out of 5 by from works good on IF of tissue samplesWe used this product for the detection of a-SMA in the mouse fibrotic liver frozen sections.
Date published: 2017-10-04
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α-Actin Antibody (1A4) is rated 4.7 out of 5 by 31.
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