Note: You clicked on an external link, which has been disabled in order to keep your shopping session open.
Immunofluorescence analysis of Phospho-Cortactin (Tyr421) was performed using 70% confluent log phase NIH/3T3 cells treated with 1.6 mM hydrogen peroxide for 1 h. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-Cortactin (Tyr421) Rabbit Polyclonal Antibody (44854G) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the untreated cells. Panel f shows the control without primary antibody. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||All, Tag, Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from the region of mouse cortactin that contains tyrosine 421.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 50% glycerol, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 2 publications below|
Cortactin is an 80/85 kDa cytoskeleton protein that facilitates assembly of cortical actin. Cortactin is widely expressed in most adherent cells and is a prominent substrate of protein-tyrosine kinase Src in vivo and in vitro. The protein sequence of cortactin contains multiple amino-terminal tandem repeats of a unique 37-amino acid sequence, which associates with actin, and a Src homology 3 (SH3) domain at the carboxyl terminus. Between the repeat and the SH3 domain, there is an alpha-helical structure followed by a proline-rich region.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Mouse||Not Cited||Crk-associated substrate tyrosine phosphorylation sites are critical for invasion and metastasis of SRC-transformed cells.||Brábek J,Constancio SS,Siesser PF,Shin NY,Pozzi A,Hanks SK||Molecular cancer research : MCR (3:307)||2005|
|Cortactin tyrosine phosphorylation requires Rac1 activity and association with the cortical actin cytoskeleton.||Head JA,Jiang D,Li M,Zorn LJ,Schaefer EM,Parsons JT,Weed SA||Molecular biology of the cell (14:3216)||2003|
1110020L01Rik; amplaxin; cortactin; CTTN; EMS1; ems1 sequence (mammary tumor and squamous cell carcinoma-associated (p80/85 src substrate); FLJ34459; mammary tumor and squamous cell carcinoma associated (p80/85 src substrate); oncogene EMS1; src substrate cortactin; SRC8
1110020L01Rik; CTTN; EMS1