Anti-alpha smooth muscle Actin antibody (ab5694)
Key features and details
- Rabbit polyclonal to alpha smooth muscle Actin
- Suitable for: WB, IHC-P
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-alpha smooth muscle Actin antibody
See all alpha smooth muscle Actin primary antibodies -
Description
Rabbit polyclonal to alpha smooth muscle Actin -
Host species
Rabbit -
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. The other actins, such as beta- and gamma-cytoplasmic, striated muscle and myocardium are not stained by this alpha smooth muscle Actin antibody.
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Tested Applications & Species
See all applications and species dataApplication Species IHC-P MouseHumanWB MouseHuman
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Immunogen
Synthetic peptide corresponding to Human alpha smooth muscle Actin (N terminal).
Database link: P62736 -
Positive control
- WB: HEK-293, A431, HeLa, Jurkat and NIH/3T3 whole cell lysate. Mouse heart tissue homogenate. HeLa nuclear lysate. IHC-P: PACT-sRIMS-cleared virgin and lactating mammary glands. Mouse intestine and pancreas tissue. Mouse aortic regurgitation. Mouse tonsil tissue.
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General notes
Actins are highly conserved proteins expressed in all eucaryotic cells. Actin filaments form part of the cytoskeleton and play essential roles in regulating cell shape and movement. Six distinct actin isotypes have been identified in mammalian cells. Each is encoded by a separated gene and is expressed in a developmentally regulated and tissue-specific manner, alpha and beta cytoplasmic actins are expressed in a wide variety of cells; whereas, expression of alpha skeletal, alpha cardiac, alpha vascular and gamma enteric actins are more restricted to specialized muscle cell type. Smooth muscle alpha actin is of further interest because it is one of a few genes whose expression is relatively restricted to vascular smooth muscle cells. Further more, expression of smooth muscle alpha actin is regulated by hormones, cell proliferation and altered by pathological conditions including oncogenic transformation and atherosclerosis.
Abcam is committed to meeting high standards of manufacturing and has decided to discontinue this product once the stock runs out as we are unable to secure its future high-quality supply. We suggest ab124964, ab150301 or ab7817 as possible replacements. We are sorry for any inconvenience this may cause.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Western blot - Anti-alpha smooth muscle Actin antibody (ab5694)Lanes 1-3 : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml
Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate - overexpressing alpha-Actin
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) cell lysate
Lanes 3 & 5 : Mouse heart tissue homogenate
Lane 4 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
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 kDaIncubated with the primary antibody at 4°C overnight.
Incubated with the secondary antibody at room temperature for 1 hour.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694) 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/.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.
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694) 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/.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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694) 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/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)
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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 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.
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Western blot - Anti-alpha smooth muscle Actin antibody (ab5694)All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) nuclear cell lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa Fluor anti-rabbit at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Additional bands at: 30 kDa, 35 kDa, 37 kDa, 50 kDa, 75 kDa. We are unsure as to the identity of these extra bands.
Please note that ab5694 does not appear to be specific to smooth muscle.
