使用敲除细胞株进行验证RabMAb

Anti-Cortactin抗体[EP1922Y] (ab81208)

概述

性能

应用

Our Abpromise guarantee covers the use of ab81208 in the following tested applications.

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

应用 Ab评论 说明
WB 1/50000 - 1/100000. Predicted molecular weight: 62 kDa.
IHC-P 1/100 - 1/250.
ICC/IF 1/1000.
Flow Cyt 1/10. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP 1/40.

靶标

  • 功能Contributes to the organization of the actin cytoskeleton and cell structure. Plays a role in the regulation of cell migration. Plays a role in the invasiveness of cancer cells, and the formation of metastases.
  • 序列相似性Contains 7 cortactin repeats.
    Contains 1 SH3 domain.
  • 结构域The SH3 motif may mediate binding to the cytoskeleton.
  • 翻译后修饰Tyrosine phosphorylation in transformed cells may contribute to cellular growth regulation and transformation.
  • 细胞定位Cytoplasm > cytoskeleton. Cell projection > lamellipodium. Cell projection > ruffle. Associated with membrane ruffles and lamellipodia.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Amplaxin antibody
    • CTTN antibody
    • EMS 1 antibody
    • EMS1 antibody
    • FLJ34459 antibody
    • Mammary tumor and squamous cell carcinoma associated antibody
    • Oncogene EMS1 antibody
    • p80/85 src substrate antibody
    • Src substrate cortactin antibody
    • SRC8_HUMAN antibody
    see all

Anti-Cortactin antibody [EP1922Y] 图像



  • Predicted band size : 62 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Cortactin knockout HAP1 cell lysate (20 µg)
    Lane 3: NIH/3T3 cell lysate (20 µg)
    Lane 4: A431 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab81208 observed at 62 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab81208 was shown to specifically react with Cortactin when Cortactin knockout samples were used. Wild-type and Cortactin knockout samples were subjected to SDS-PAGE. ab81208 and ab8245 (loading control to GAPDH) were both diluted 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab81208 staining Cortactin in wild-type HAP1 cells (top panel) and Cortactin knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab81208 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Anti-Cortactin antibody [EP1922Y] (ab81208) at 1/100000 dilution + HeLa cell lysate at 10 µg

    Secondary
    HRP labelled Goat anti-Rabbit at 1/2000 dilution

    Predicted band size : 62 kDa
  • Immunohistochemical staining of paraffin-embedded human breast carcinoma using 1/100 ab81208.
  • Immunofluorescent staining of MCF7 cells using 1/100 ab81208
  • Overlay histogram showing HeLa cells stained with ab81208 (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 (ab81208, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-Cortactin antibody [EP1922Y] (ab81208)参考文献

This product has been referenced in:
  • Motonishi S  et al. Sirtuin1 Maintains Actin Cytoskeleton by Deacetylation of Cortactin in Injured Podocytes. J Am Soc Nephrol 26:1939-59 (2015). Read more (PubMed: 25424328) »
  • Kaur S  et al. An unmet actin requirement explains the mitotic inhibition of clathrin-mediated endocytosis. Elife 3:e00829 (2014). ICC/IF ; Human . Read more (PubMed: 24550251) »

See all 4 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 20°C
Sample Human Cell (Ovarian carcinoma (A2780))
Specification Ovarian carcinoma (A2780)
Permeabilization Yes - 0.1% Triton-x100
Fixative Paraformaldehyde
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提交于 Aug 18 2014

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