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Anti-alpha smooth muscle Actin antibody (ab5694) is a rabbit polyclonal antibody detecting alpha smooth muscle Actin in Western Blot, IHC-P. Suitable for Human, Mouse.



- Over 3000 publications

- Trusted since 2003



Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694), expandable thumbnail
  • Western blot - Anti-alpha smooth muscle Actin antibody (AB5694), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (AB5694), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.4
Preservative: 0.05% Sodium azide
Constituents: 99.95% PBS

Form
Liquid
Clonality
Polyclonal

Immunogen

  • Synthetic Peptide within Human ACTA2. The exact immunogen used to generate this antibody is proprietary information. Database link P62736

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Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-P
Human
Tested
Tested
Mouse
Tested
Tested

Tested
Tested

Species
Human
Dilution info
0.5-2 µg/mL
Notes

-

Species
Mouse
Dilution info
0.5-2 µg/mL
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/50 - 1/200
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Species
Human
Dilution info
1/50 - 1/200
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

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Target data

Function

Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.

Alternative names

Recommended products

Anti-alpha smooth muscle Actin antibody (ab5694) is a rabbit polyclonal antibody detecting alpha smooth muscle Actin in Western Blot, IHC-P. Suitable for Human, Mouse.



- Over 3000 publications

- Trusted since 2003


Key facts

Isotype
IgG
Form
Liquid
Clonality
Polyclonal
Immunogen
  • Synthetic Peptide within Human ACTA2. The exact immunogen used to generate this antibody is proprietary information. Database link P62736
Purification technique
Affinity purification Immunogen
Specificity

Alpha smooth muscle actin antibody (ab5694) stains smooth muscle cells in vessel walls, gut wall, and myometrium. Myoepithelial cells in breast and salivary gland are also stained. ab5694 reacts with tumors arising from smooth muscles and myoepithelial cells.

Concentration
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Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Anti-alpha smooth muscle Actin antibody (ab5694) is a rabbit polyclonal antibody and is validated for use in IHC-P and WB.

Anti-alpha smooth muscle Actin antibody (ab5694) was first used in a scientific publication in 1985 and has been cited over 3004 times in peer reviewed journals. It's performance in Western blot, immunofluorescence and IHC in human, mouse and rat samples is trusted by the scientific community.

Abcam's high quality validation processes ensure Anti-alpha smooth muscle Actin antibody (ab5694) has high sensitivity and specificity.

Anti-alpha smooth muscle Actin antibody (ab5694) has 150 independent reviews from customers.

Anti-alpha smooth muscle Actin antibody (ab5694) specifically detects alpha smooth muscle Actin (UniProt ID: P62736; Molecular weight: 42kDa) and is sold in 100 µg selling sizes.

Alpha-Smooth Muscle Actin (α-SMA) is a critical protein involved in smooth muscle contraction, encoded by the ACTA2 gene. Predominantly found in vascular smooth muscle cells, α-SMA plays a vital role in fibrogenesis and wound healing. It is often used as a marker for myofibroblast formation, which are specialized cells that contribute to tissue repair and fibrosis. The expression of α-SMA is associated with the activation of myofibroblasts, making it an important indicator in studies of fibrosis and other related conditions.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

Alpha smooth muscle actin (α-SMA) also known as ACTA2 is an actin isoform with a specific role in the contractile function of smooth muscle cells. The molecular weight of α-SMA is approximately 42 kDa. This protein is expressed widely in vascular smooth muscle cells the peritoneal lining and myofibroblasts. It often serves as a marker for these cell types. The expression of α-SMA is critical for the mechanical attributes of cells contributing to the rigidity and contractility of tissues where it is present.

Biological function summary

Alpha smooth muscle actin aids in maintaining the structural integrity of tissues by forming part of the actin cytoskeleton an important element in cellular support. It exists in high concentration in stress fibers contributing to cellular movements and shape maintenance. These actions are essential in various dynamic cellular processes such as cell migration and adhesion. Alpha smooth muscle actin does not typically form complexes but it associates with other components of the actin cytoskeleton to ensure cell stability and function.

Pathways

Alpha smooth muscle actin functions significantly within the TGF-beta signaling pathway which influences cell proliferation differentiation and apoptosis. It interacts closely with proteins such as myosin to facilitate cellular contractility and motility. Additionally α-SMA plays a part in the RhoA/Rho kinase (ROCK) pathway connecting with regulators of actin filament organization. These pathways are essential for modulation of smooth muscle contraction and actin filament assembly contributing to vascular development and wound healing.

Associated diseases and disorders

Alpha smooth muscle actin is importantly involved in fibrotic diseases and vascular diseases like arteriosclerosis. Increased expression of α-SMA is often observed in fibrotic tissues contributing to pathogenesis due to excessive extracellular matrix deposition. In the context of vascular disorders overexpression of α-SMA can lead to abnormal vascular remodeling often seen in diseases like hypertension. Interactions with proteins such as fibronectin and collagens facilitate these pathological changes highlighting the importance of α-SMA in disease development.

Product promise

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8 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694), expandable thumbnail
    This image is courtesy of a customer review submitted by Cameron Johnstone.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of 10% formalin-fixed Human transitional cell carcinoma of the kidney. Stained with ab5694 at 1/150 dilution. Secondary antibody used was goat anti-rabbit Alexa-568® at 1/500 dilution. Blocking was done with 5% serum for 1 hour at 25°C. The sample was incubated with the primary antibody for 1hour at 25°C with 5% normal goat serum. Antigen retrieval method was heat mediated with EDTA pH 8.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694), expandable thumbnail
    Image from Lee H. et al., BMC Dev Biol. 2014 Dec 21;14:48. Fig 2Bdoi: 10.1186/s12861-014-0048-3 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

    Pancreatic vessel imaging in the intact adult mouse pancreas. In adult mouse tissues (12 weeks old), imaging was performed after CLARITY. Three-dimensional projection clarified mouse pancreas with capillary immunostained for α-smooth muscle actin (green). Scale bar, 200 μm.

    From a study, that aimed to improve the original CLARITY procedure and developed specific CLARITY protocols for various intact organs.

  • Western blot - Anti-alpha smooth muscle Actin antibody (ab5694), expandable thumbnail

    Western blot - Anti-alpha smooth muscle Actin antibody (ab5694)

    Incubated with the primary antibody at 4°C overnight.

    Incubated with the secondary antibody at room temperature for 1 hour.

    Lanes 1 - 3: Western blot - Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/mL

    Lanes 4 - 5: Western blot - Anti-alpha smooth muscle Actin antibody (ab5694)

    Lane 1: HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate - overexpressing alpha-Actin at 20 µg

    Lane 2: NIH/3T3 (Mouse embryonic fibroblast cell line) cell lysate at 20 µg

    Lanes 3 and 5: Mouse heart tissue homogenate at 20 µg

    Lane 4: NIH/3T3 cell lysate at 20 µg

    Secondary

    All lanes: Fluor 750-conjugated goat anti-rabbit IgG (H+L) at 1/12500 dilution

    Predicted band size: 42 kDa

    Observed band size: 42 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694), expandable thumbnail
    Image from Lloyd-Lewis B. et al., Breast Cancer Res. 2016 18: 127. Fig 2.; doi: 10.1186/s13058-016-0754-9. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

    Passive CLARITY technique (PACT)-sorbitol refractive index matching solution (sRIMS) clearing and 3D imaging of virgin and lactating mouse mammary tissue. a PACT-sRIMS tissue clearing and immunostaining protocol and timeline. Three-dimensional confocal imaging of PACT-sRIMS-cleared virgin (b) and lactating (c) mammary glands immunostained with basal cell markers (K5 and smooth muscle actin (stained with ab5694)) and luminal cell markers (K8 and E-cadherin (E-CAD)). Main image shows the maximum intensity projection of the entire image sequence, with thin optical slices (1 μm) and their depth (z value) relative to the first image in the image sequence.

    From a paper comparing imaging of intact virgin and lactating mammary glands using 3D imaging of solvent-cleared organs, see deep brain (seeDB), clear unobstructed brain imaging cocktails (CUBIC) and passive clarity technique.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

    This picture shows formalin-fixed, paraffin embedded mouse intestine and mesentery, the optimal dilution is 1:1600 to 1:3200, incubation overnight at 4oC, counterstained with Hematoxylin.

    This image was kindly supplied as part of the review by JQ Zhang.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694), expandable thumbnail
    Freytsis, M. et al PLoS One. 2018 Jan 5;13(1):e0190623. doi: 10.1371/journal.pone.0190623. eCollection 2018 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

    Representative histology of aortic valve leaflets from aged mice demonstrates changes in pRb cKO AoV

    A) Masson's trichrome showing reduced collagen staining (blue) in leaflet from pRb cKO mouse with aortic regurgitation (AR). B) Movat pentachrome showing more diffuse collage staining (yellow) in fibrosa, but normal proteoglycan staining (blue) in the spongiosa layer of the leaflet from pRb cKO with AR. C) Immunohistochemistry for α-SMA, demonstrating presence of activated myofibroblasts throughout leaflets of pRb cKO mouse with and without AR. Scale bar is 50μm.

    Alpha smooth muscle Actin is detected witn ab5694 at 1/1000 dilution.

    (From Figure 2 of Freytsis et al)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling alpha smooth muscle Actin with ab5694 at a dilution of 1/1000. Heat mediated antigen retrieval was performed for 35 minutes followed by cooling for 20 minutes. Sections were incubated with the primary antibody for 1 hour followed by incubation with a biotinylated secondary antibody (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) for 30 minutes then HRP-Streptavidin for 30 minutes. Developed using DAB chromogen substrate (5-10 minutes). Counter stained with hematoxylin.

    Magnification: left - 10X, right - 40X.

  • Western blot - Anti-alpha smooth muscle Actin antibody (ab5694), expandable thumbnail

    Western blot - Anti-alpha smooth muscle Actin antibody (ab5694)

    Please note that ab5694 does not appear to be specific to smooth muscle.

    All lanes: Western blot - Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/mL

    Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear cell lysate at 20 µg

    Lane 2: HeLa whole cell lysate at 20 µg

    Lane 3: A431 (Human epidermoid carcinoma cell line) cell lysate at 20 µg

    Lane 4: Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate at 20 µg

    Lane 5: HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 20 µg

    Secondary

    All lanes: Alexa Fluor anti-rabbit at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 42 kDa

    Observed band size: 30 kDa, 35 kDa, 37 kDa, 42 kDa, 50 kDa, 75 kDa

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