Anti-alpha Actinin 抗体 [EP2527Y] (ab68194)

製品の概要

  • 製品名
    Anti-alpha Actinin antibody [EP2527Y]
    alpha Actinin 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP2527Y] to alpha Actinin
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: WB, IP, Flow Cyt, IHC-Pmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    A synthetic peptide corresponding to residues on the N-terminus of human alpha Actinin

  • ポジティブ・コントロール
    • NIH 3T3 lysate, HeLa lysate and C6 lysate
  • 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

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab68194 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 1/2000 - 1/10000. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).
IP 1/40.
Flow Cyt 1/100 - 1/1000.

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

IHC-P 1/100.

ターゲット情報

  • 機能
    F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein.
  • 配列類似性
    Belongs to the alpha-actinin family.
    Contains 1 actin-binding domain.
    Contains 2 CH (calponin-homology) domains.
    Contains 2 EF-hand domains.
    Contains 4 spectrin repeats.
  • 細胞内局在
    Cytoplasm > cytoskeleton. Cytoplasm > myofibril > sarcomere > Z line. Colocalizes with MYOZ2 and PPP3CA at the Z-line of heart and skeletal muscle.
  • Information by UniProt
  • 参照データベース
  • 別名
    • actinin 1 smooth muscle antibody
    • Actinin alpha 1 antibody
    • actinin, alpha 1 antibody
    • ACTN 1 antibody
    • Actn1 antibody
    • ACTN1_HUMAN antibody
    • Alpha Actinin 1 antibody
    • Alpha actinin cytoskeletal isoform antibody
    • Alpha-actinin cytoskeletal isoform antibody
    • Alpha-actinin-1 antibody
    • BDPLT15 antibody
    • F actin cross linking protein antibody
    • F-actin cross-linking protein antibody
    • FLJ40884 antibody
    • FLJ54432 antibody
    • Non muscle alpha actinin 1 antibody
    • Non-muscle alpha-actinin-1 antibody
    see all

画像

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Alpha Actinin knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: NIH 3T3 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab68194 observed at 103 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab68194 was shown to specifically react with alpha Actinin in wild-type HAP1 cells as signal was lost in alpha Actinin knockout cells. Wild-type and alpha Actinin knockout samples were subjected to SDS-PAGE. ab68194 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-alpha Actinin antibody [EP2527Y] (ab68194) at 1/10000 dilution

    Lane 1 : NIH 3T3 lysate
    Lane 2 : HeLa lysate
    Lane 3 : C6 lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 103 kDa
    Observed band size: 103 kDa

  • ab68194 staining alpha Actinin in rat kidney tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 30% BSA for 10 minutes at 37°C; antigen retrieval was by heat mediation in EDTA. Samples were incubated with primary antibody (1/100 in PBS) for 16 hours at 4°C. ab80436 EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit was used.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab68194 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab68194, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG; H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

参考文献

ab68194 has not yet been referenced specifically in any publications.

レビューと Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 30% · Temperature: 37°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA
Sample
Rat Tissue sections (kidney)
Specification
kidney
Permeabilization
No
Fixative
Formaldehyde
Username

Prof. Kadir Demircan

Verified customer

投稿 Oct 10 2014

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

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