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RabMAb

Anti-alpha smooth muscle Actin 抗体 [E184] (ab32575)

製品の概要

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32575 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-FrFl Use at an assay dependent concentration. PubMed: 24647450
IHC-Fr 1/100.
IF Use at an assay dependent concentration. PubMed: 28138565
WB 1/1000 - 1/5000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human alpha smooth muscle Actin peptide (ab211918).
IHC-P 1/200 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol). Internal test showed non-specific staining on mouse kidney, mouse stomach, rat kidney and rat stomach.

Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/500.
  • 追加情報
    Is unsuitable for IP.
  • 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

    Anti-alpha smooth muscle Actin antibody [E184] 画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human smooth muscle tissue labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labelling alpha smooth muscle Actin (green) with purified ab32575 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by ab150120 AlexaFluor®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).

      For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).  

    • All lanes : Anti-alpha smooth muscle Actin antibody [E184] (ab32575) at 1/5000 dilution (purified)

      Lane 1 : A431 whole cell lysate
      Lane 2 : HeLa whole cell lysate
      Lane 3 : MCF-7 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 42 kDa
      Observed band size : 42 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • Flow Cytometry analysis of HeLa cells labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/20 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • All lanes : Anti-alpha smooth muscle Actin antibody [E184] (ab32575) at 1/5000 dilution (purified)

      Lane 1 : C6 whole cell lysate
      Lane 2 : RAW264.7 whole cell lysate
      Lane 3 : PC-12 whole cell lysate
      Lane 4 : NIH/3T3 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 42 kDa
      Observed band size : 42 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse smooth muscle tissue labelling alpha smooth muscle Actin with purified ab32575 at a dilution of 1/200. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • Unpurified ab32575 staining alpha smooth muscle actin in human kidney tumour sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 10% serum for 20 minutes at 21°C. Samples were incubated with primary antibody (1/100 in TBS + 2% BSA + 0.02% sodium azide) for 1 hour at 21°C. An undiluted HRP-conjugated Goat anti-rabbit polyclonal was used as the secondary antibody.

    Anti-alpha smooth muscle Actin antibody [E184] (ab32575) 使用論文

    This product has been referenced in:
    • Zheng L  et al. Baicalin ameliorates renal fibrosis via inhibition of transforming growth factor ß1 production and downstream signal transduction. Mol Med Rep 15:1702-1712 (2017). IHC-P ; Rat . Read more (PubMed: 28260014) »
    • Wang Q  et al. HMGB1 induces lung fibroblast to myofibroblast differentiation through NF-?B-mediated TGF-ß1 release. Mol Med Rep 15:3062-3068 (2017). Read more (PubMed: 28339089) »

    See all 91 Publications for this product

    Product Wall

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Heart)
    Permeabilization
    Yes - 0.2% triton TX-100
    Specification
    Heart
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 Mar 04 2016

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
    Permeabilization
    Yes - Wash Buffer from Dako with Tween
    Specification
    Liver
    Fixative
    Paraformaldehyde
    Username

    Mr. Rudolf Jung

    Verified customer

    投稿 Dec 21 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer ph 9,0
    Permeabilization
    Yes - Wash Buffer from Dako with Tween
    Specification
    Skin
    Fixative
    Paraformaldehyde
    Username

    Mr. Rudolf Jung

    Verified customer

    投稿 Dec 21 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Lung)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA
    Permeabilization
    Yes - 0.5% tritonX-100
    Specification
    Lung
    Blocking step
    invitrogen kit blocking serum as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 Sep 09 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: sodium citrate pH 6.0
    Sample
    Mouse Tissue sections (Skin wound)
    Specification
    Skin wound
    Permeabilization
    Yes - 0.05% Tween 20
    Fixative
    10% formalin
    Username

    Abcam user community

    Verified customer

    投稿 Oct 27 2014

    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native)
    Sample
    Mouse Tissue lysate - whole (liver)
    Specification
    liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    投稿 May 05 2014

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Sodium Citrate Buffer pH 6.0
    Sample
    Mouse Tissue sections (liver)
    Specification
    liver
    Permeabilization
    Yes - Sodium Citrate Buffer pH 6.0
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 May 02 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 1.5% · Temperature: 21°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: antigen retrieval
    Sample
    Mouse Tissue sections (Heart at day 0 and day 7 post-MI)
    Specification
    Heart at day 0 and day 7 post-MI
    Permeabilization
    No
    Fixative
    10% zinc formalin
    Username

    Dr. Merry Lindsey

    Verified customer

    投稿 Sep 20 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris Gel)
    Sample
    Mouse Tissue lysate - whole (heart)
    Specification
    heart
    Treatment
    day 0 and day 7 after myocardial infarction
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Username

    Dr. Merry Lindsey

    Verified customer

    投稿 Sep 20 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated
    Sample
    Mouse Tissue sections (blood vessels in the soft tissue)
    Specification
    blood vessels in the soft tissue
    Permeabilization
    Yes - 0.1% saponin
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 Apr 29 2013

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