使用敲除细胞株进行验证

Anti-Cortactin抗体[4F11] (ab33333)

概述

  • 产品名称Anti-Cortactin抗体[4F11]
    参阅全部 Cortactin 一抗
  • 描述
    小鼠单克隆抗体[4F11] to Cortactin
  • 特异性The clone number has been updated from (0.T.21) to (4F11) both clone numbers name the same antibody clone.
  • 经测试应用适用于: ICC/IF, WB, IP, ICCmore details
  • 种属反应性
    与反应: Mouse, Rat, Chicken, Hamster, Cow, Human
  • 免疫原

    Mixture of affinity purified tyrosine phosphoproteins from chick embryo fibroblasts expressing activated pp60 src.

  • 阳性对照
    • WB: Mouse 3T3 cell lysate and chicken fibroblast src transformed cell lysate. ICC/IF: CTTN-HAP1 wild-type and knockout cells

性能

应用

Our Abpromise guarantee covers the use of ab33333 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 61 kDa. Use 20 µg per lane of 3T3 cell lysate as a positive control for mini gels.
Note: ab33333 recognizes 80kD and 85kD proteins in avian cells, but it recognizes an 80kD protein in rodent and human cells.
IP Use at 10 µg/mg of lysate.
ICC Use a concentration of 10 µg/ml.

靶标

Anti-Cortactin antibody [4F11] 图像

  • ab33333 staining Cortactin in wild-type HAP1 cells (top panel) and CTTN knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), 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 ab33333 at 1μg/ml and ab7291 at 1μg/ml 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) and goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo colour red). Nuclear DNA was labelled in blue with DAPI.

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

  • Anti-Cortactin antibody [4F11] (ab33333) at 1/1000 dilution + Bovine aortic endothelial whole cell lysates at 10 µg

    Secondary
    Goat anti-mouse polyclonal HRP conjugate at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 61 kDa
    Additional bands at : 80 kDa (possible post-translational modification).

    Exposure time : 5 seconds

    Image is courtesy of an anonymous AbReview

    Blocking was with 5% milk for 1 hour at 20°C.

    See Abreview

  • ab33333 staining Cortactin (red) in Human palladin-activated fibroblasts by Immunocytochemistry/ Immunofluorescence.

    Cells were fixed with paraformaldehyde and permeabilized with 0.2% Triton X-100. Samples were incubated with primary antibody (1/500 in diluent) before incubation with a Rhodamine Red IgG (1/100).

Anti-Cortactin antibody [4F11] (ab33333)参考文献

This product has been referenced in:
  • Weidmann MD  et al. Mena(INV) dysregulates cortactin phosphorylation to promote invadopodium maturation. Sci Rep 6:36142 (2016). Read more (PubMed: 27824079) »
  • Myers RB  et al. The matricellular protein CCN5 regulates podosome function via interaction with integrin avß 3. J Cell Commun Signal 8:135-46 (2014). Rat . Read more (PubMed: 24488697) »

See all 11 Publications for this product

Product Wall

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (COS7 cell)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification COS7 cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Oct 16 2015

Application Western blot
Sample Dog Cell lysate - whole cell (Madin-Darby Canine Kidney (MDCK) epithelial cell)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification Madin-Darby Canine Kidney (MDCK) epithelial cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Sep 29 2015

Application Western blot
Sample Cow Cell lysate - whole cell (Bovine aortic endothelial cell)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification Bovine aortic endothelial cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Sep 25 2015

Application Western blot
Sample Human Cell lysate - whole cell (HEK293T cells)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification HEK293T cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Jun 19 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 15 minute(s) · Concentration: 0.5% · Temperature: 20°C
Sample Mouse Cell (J774 macrophage)
Specification J774 macrophage
Permeabilization Yes - 0.05% saponin
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 May 23 2014

Thank you for contacting us. I am sorry to hear that this antibody is not providing satisfactory results.

Would you be able to provide the lot number and order number?

It may be possible that the ˜100 kDa band corresponds to the pho...

Read More

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