製品の概要

  • 製品名
    Anti-alpha smooth muscle Actin antibody
    alpha smooth muscle Actin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to alpha smooth muscle Actin
  • 由来種
    Rabbit
  • 特異性
    This antibody stains smooth muscle cells in vessel walls, gut wall, and myometrium. Myoepithelial cells in breast and salivary gland are also stained. It 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 antibody.
  • アプリケーション
    適用あり: IHC-FoFr, ICC, ICC/IF, WB, ELISA, IHC-P, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Rat, Chicken, Guinea pig, Cow, Dog, Human, Pig
  • 免疫原

    Synthetic peptide corresponding to Human alpha smooth muscle Actin. This antibody was raised against a synthetic peptide corresponding to N-terminus of actin from human smooth muscle.

  • 特記事項

    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 recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab5694 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
IHC-FoFr Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
ICC/IF 1/100.
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 42 kDa.
ELISA Use a concentration of 0.1 - 1 µg/ml.
IHC-P 1/50 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr 1/200. PubMed: 18559614Fix with acetone.

ターゲット情報

  • 機能
    Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
  • 関連疾患
    Defects in ACTA2 are the cause of aortic aneurysm familial thoracic type 6 (AAT6) [MIM:611788]. AATs are characterized by permanent dilation of the thoracic aorta usually due to degenerative changes in the aortic wall. They are primarily associated with a characteristic histologic appearance known as 'medial necrosis' or 'Erdheim cystic medial necrosis' in which there is degeneration and fragmentation of elastic fibers, loss of smooth muscle cells, and an accumulation of basophilic ground substance.
  • 配列類似性
    Belongs to the actin family.
  • 細胞内局在
    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • 参照データベース
  • 別名
    • a actin antibody
    • AAT6 antibody
    • ACTA_HUMAN antibody
    • ACTA2 antibody
    • Actin alpha 2 smooth muscle aorta antibody
    • Actin aortic smooth muscle antibody
    • Actin, aortic smooth muscle antibody
    • ACTSA antibody
    • ACTVS antibody
    • Alpha 2 actin antibody
    • Alpha actin 2 antibody
    • Alpha cardiac actin antibody
    • alpha sma antibody
    • Alpha-actin-2 antibody
    • Cell growth inhibiting gene 46 protein antibody
    • Cell growth-inhibiting gene 46 protein antibody
    • GIG46 antibody
    • Growth inhibiting gene 46 antibody
    • MYMY5 antibody
    see all

画像

  • Effects of the ROCK1 inhibition on pancreatic cancer cells and cancer-associated fibroblasts

    Fluorescence microscopic analysis of fasudil treated, co-cultured pancreatic cancer cells and cancer-associated fibroblasts. Cells were treated with fasudil for 48 hours and then were stained for α-SMA (red), Collagen I (green), and DNA (blue).

    Alpha smooth muscle Actin (α-SMA) is detected ising ab5694 in 5% formaldehyde-fixed cells.

    (From Figure 3C of Watcott et al)

  • 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)

  • 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.

  • ab5694 at 1/500 staining rat myofibroblast cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed and blocked with 5% serum prior to incubation with the antibody for 2 hours. A FITC conjugated goat anti-rabbit IgG was used as the secondary. Nuclei were counterstained with propidium iodide.

    See Abreview

  • Lanes 1-3 : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml


    Lane 1 : HEK293 cell lysate - overexpressing alpha-Actin
    Lanes 2 & 4 : 3T3 cell lysate
    Lanes 3 & 5 : Mouse heart tissue homogenate

    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 kDa



    Incubated with the primary antibody at 4°C overnight.

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

  • Ab5694 positively staining smooth muscle cells in blood vessels and myoepithelial cells in the frozen tissue of cancerous human mammary gland (pink) at 1/100 dilution. Secondary: CY5 conjugated goat anti rabbit (1/100). Co immunostaining of glandular cell cytokeratin can be seen stained by FITC (green). Auto fluorescent erythrocytes that are present within blood vessels are shown (red), whilst the DAPI counter stain may clearly be seen staining nuclei (blue).

    This image is courtesy of an Abreview submitted by on 22 August 2005. We do not have any further information relating to this image.

    See Abreview

  • ab5694 staining Human fetal heart cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37°C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37°C. A  Alexa Fluor® 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.

    See Abreview

  • 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.

  • Immunohistochemistry (Formalin-fixed paraffin-embedded sections) analysis of skeletal muscle tissue (left) incubated with ab5694 at 1/100 at room temperature for 1 hour showing no specific staining. Right - human tonsil tissue secondary only control.

    Heat mediated antigen retrieval was performed for 35 minutes followed by cooling for 20 minutes. A biotinylated secondary antibody was used for 30 minutes followed by incubation with HRP-Streptavidin for 30 minutes. Developed using DAB chromogen substrate (5-10 minutes). Counter stained with hematoxylin. Magnification 10X.

  • Human Leiomyoma stained with ab5694.
  • All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml

    Lane 1 : HeLa Nuclear
    Lane 2 : HeLa whole cell
    Lane 3 : A431 cell lysate
    Lane 4 : Jurkat cell lysate
    Lane 5 : HEK293 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.
  • All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1/500 dilution

    Lane 1 : Rat2 myofibroblasts (untreated before treatment-0 days)
    Lane 2 : Rat2 myofibroblasts (untreated for 5 days)
    Lane 3 : Rat2 myofibroblasts (treated with 1ng/mL TGF beta)
    Lane 4 : Rat2 myofibroblasts (treated with 10ng/mL TGF beta)
    Lane 5 : Positive control (NIH3T3)
    Lane 6 : Negative control (MDA-MB-469 breast carcinoma cells)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Donkey anti rabbit (HRP) at 1/2500 dilution

    Performed under reducing conditions.

    Predicted band size: 42 kDa



    This image is an edited version of an image submitted courtesy of an Abreview on 20 September 2005. We do not have any further information relating to this image.

    See Abreview

  • All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1/1000 dilution

    Lanes 1-2 : Lystates prepared from pig heart tissue from normal control animals
    Lanes 3-4 : Lystates prepared from pig heart tissue from experimental animals

    Lysates/proteins at 4 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat polyclonal to rabbit IgG at 1/20000 dilution

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute

    See Abreview

  • ab5694 staining alpha smooth muscle Actin in human skin tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in Citrate pH 6.0 and then blocked with 10% serum for 1 hour at RT. The primary antibody was diluted 1/300 and incubated with sample in 2% serum for 15 hours at 4°C. A Biotin conjugated goat polyclonal to rabbit IgG was used at dilution at 1/500 as secondary antibody.

    See Abreview

  • ab5694 staining alpha smooth muscle Actin in rat lung tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
    Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then permeabilized using 0.5 Triton X-100 for 20 minutes, blocked with 1% BSA for 30 minutes at 20°C and then incubated with ab5694 at a 1/100 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 568 conjugated goat anti-rabbit polyclonal used at a 1/250 dilution.

    See Abreview

参考文献

This product has been referenced in:
  • Luo L  et al. Combination treatment of adipose-derived stem cells and adiponectin attenuates pulmonary arterial hypertension in rats by inhibiting pulmonary arterial smooth muscle cell proliferation and regulating the AMPK/BMP/Smad pathway. Int J Mol Med 41:51-60 (2018). Read more (PubMed: 29115380) »
  • Freytsis M  et al. Conditional deletion of RB1 in the Tie2 lineage leads to aortic valve regurgitation. PLoS One 13:e0190623 (2018). Read more (PubMed: 29304157) »
See all 1002 Publications for this product

レビューと Q&A

1-10 of 144 Abreviews or Q&A

Answer

I have checked the website and the datasheets of these three antibodies and it seems that only ab18147 has been tested for cross-reactivity with rabbit. The other two antibodies are not known yet if they recognize rabbit alpha smooth muscle Actin.

However, it is important to mention that according to the datasheet ab18147 recognizes the alpha-smooth muscle isoform of actin but it does not react with actin from fibroblasts (b-and g-cytoplasmic), striated muscle (a-sarcomeric), and myocardium (a-myocardial).

We have a nice range of products for detection (ie. anti-mouse HRP based detection kits and reagents), you may wish to visit this site to get some further information:

https://www.abcam.com/index.html

Read More

Answer

We do not have set expiration dates for our products. I am therefore also not aware that the information on the shelf life was changed. We indeed increased the length of our Abpromise guarantee to six month instead of 120 days.

I can confirm that most antibodies are stable and can last for anywhere from a few months to several years if stored properly, so we strongly recommend that you follow the storage instructions on the datasheet for the antibody you purchased.

These conditions will vary among our antibodies, therefore, it is important to verify the storage conditions for each of our products when you receive them. We guarantee all of our products to work for at least six months from the date of purchase when stored correctly.

For more information on antibody storage and stability, please visit our Antibody Storage Guide on our Protocols and Troubleshooting Tips page (https://www.abcam.com/index.html?pageconfig=popular_protocols).

Read More

Answer

I have some information which may be of interest to you in regards to anti-alpha smooth muscle Actin antibody (ab5694).

I have found reports of alpha-SMA expression in endothelial cells. These can be found from the links below:

http://www.ncbi.nlm.nih.gov/pubmed/19285011

http://www.ncbi.nlm.nih.gov/pubmed/10330261

http://www.ncbi.nlm.nih.gov/pubmed/15588509

It may therefore be that the band you are seeing is stillspecificfor smooth muscle but expressed in the epithelial layer.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Vascular smooth muscle cell)
Permeabilization
Yes - NP40
Specification
Vascular smooth muscle cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
Fixative
Formaldehyde

Abcam user community

Verified customer

投稿 Nov 07 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% formalin

Abcam user community

Verified customer

投稿 Oct 23 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% formalin

Abcam user community

Verified customer

投稿 Oct 23 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (artery)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate pH6
Permeabilization
No
Specification
artery
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% formalin

Abcam user community

Verified customer

投稿 Oct 22 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% formalin

Abcam user community

Verified customer

投稿 Oct 22 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (artery)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
artery
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Oct 22 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Sodium Citrate pH6
Permeabilization
No
Specification
Liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

投稿 Oct 22 2018

1-10 of 144 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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