リコンビナント
RabMAb

Anti-alpha smooth muscle Actin 抗体 [EPR5368] (ab124964)

製品の概要

  • 製品名
    Anti-alpha smooth muscle Actin antibody [EPR5368]
    alpha smooth muscle Actin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR5368] to alpha smooth muscle Actin
  • アプリケーション
    適用あり: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human alpha smooth muscle Actin aa 1-100 (N terminal).
    Database link: P62736

  • ポジティブ・コントロール
    • WB: HeLa, HEK293, U937, SV40LT-SMC, A549, C2C12 and A431 cel lysate. Mouse and rat brain and heart tissue lysates. Human heart and skeletal muscle tissue lysates. IHC-P: Human prostatic carcinoma, stomach carcinoma, tonsil, heart, skeletal muscle (exhibits vascular smooth muscle staining), normal stomach, liver, colon and ovary tissues. ICC/IF: A673 cells. Flow Cyt: Jurkat cells.
  • 特記事項

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

関連製品

アプリケーション

Our Abpromise guarantee covers the use of ab124964 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
WB 1/10000 - 1/50000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).

For unpurified use at 1/1000.

IHC-P 1/1000 - 1/2500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Flow Cyt 1/10 - 1/100.

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

ICC/IF 1/250 - 1/500.

 

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovary tissue labelling alpha smooth muscle Actin with unpurified ab124964 at 1/1000 dilution.

  • All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/10000 dilution

    Lane 1 : Human lung lysate
    Lane 2 : A431 cell lysate
    Lane 3 : A549 cell lysate
    Lane 4 : NIH3T3 cell lysate
    Lane 5 : SV40-LT cell lysate
    Lane 6 : C2C12 cell lysate

    Lysates/proteins at 20 µg per lane.

    Developed using the ECL technique

    Predicted band size : 42 kDa
    Observed band size : 42 kDa


    Exposure time : 10 seconds

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% milk before being incubated with ab124964 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

  • All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/10000 dilution (purified)

    Lane 1 : HeLa cell lysate
    Lane 2 : HEK293 cell lysate
    Lane 3 : U937 cell lysate
    Lane 4 : A431 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 buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/50000 dilution (purified)

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue 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 buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/1000 dilution (unpurified)

    Lane 1 : Human heart tissue lysate - total protein (ab29431)
    Lane 2 : Heart (Mouse) Tissue Lysate
    Lane 3 : Heart (Rat) Tissue Lysate
    Lane 4 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 5 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 6 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 7 : SV40LT-SMC (Rat aorta smooth muscle) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 42 kDa
    Additional bands at : 42 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 30 seconds
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling alpha smooth muscl Actin with purified ab124964 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human heart tissue labelling alpha smooth muscle actin with unpurified ab124964 at 1/1000 dilution. Note positive staining on smooth muscle cells but negative on striated muscle cells. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human liver vessels tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human colon smooth muscle tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic carcinoma smooth muscles tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis showing vascular smooth muscle staining in skeletal muscle tissue using  alpha smooth muscle Actin with unpurified ab124964. Note positive staining on smooth muscle cells but negative on striated muscle cells.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach carcinoma smooth muscles tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human tonsil vessels tissue labelling alpha smooth muscle Actin with unpurified ab124964.

  • Immunocytochemistry/Immunofluorescence analysis of A673 cells labelling alpha smooth muscle Actin with purified ab124964 at 1/300. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

    Control: primary antibody (1/300) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).

  • Flow cytometry analysis of Jurkat cells labelling alpha smooth muscle Actin with purified ab124964 at 1/30 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

参考文献

This product has been referenced in:
  • Nakaya M  et al. Cardiac myofibroblast engulfment of dead cells facilitates recovery after myocardial infarction. J Clin Invest 127:383-401 (2017). IHC-P ; Human . Read more (PubMed: 27918308) »
  • Xu JG  et al. TGF-ß1-induced differentiation of SHED into functional smooth muscle cells. Stem Cell Res Ther 8:10 (2017). Read more (PubMed: 28114966) »

See all 14 Publications for this product

レビューと Q&A

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 post-MI
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 Nov 28 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 20 minute(s) · Concentration: 1.5% · Temperature: 21°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: antigen retrieval
Sample
Mouse Tissue sections (Heart-LV at day 0 and day 7 post-MI)
Specification
Heart-LV at day 0 and day 7 post-MI
Permeabilization
No
Fixative
10% zinc formalin
Username

Dr. Merry Lindsey

Verified customer

投稿 Sep 24 2013

I can confirm that both ab52897 and ab124964 are rabbit IgG monoclonals.

We have several suitable isotype controls available including the following. These have both been tested in IHC-P. I can recommend to review the online datasheets for f...

Read More

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

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