ANTI-CORTACTIN (P80/85), CLONE 5G3.2

Application

This mouse monoclonal Anti-Cortactin (p80/85), clone 5G3.2 Antibody, Cat. No. MABT524 is validated for use in Western Blotting for the detection of Cortactin.

Research CategoryCell Structure

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Src substrate cortactin (UniProt Q14247; also known as Amplaxin, Oncogene EMS1) is encoded by the CTTN (also known as EMS1) gene (Gene ID 2017) in human. Cortactin is a cytosolic protein that promotes polymerization and rearrangement of the actin cytoskeleton, especially the actin cortex around the cellular periphery. Cortactin is activated via phosphorylation by upstream kinases in response to extracellular stimuli. Src is known to phosphorylate cortactin on Tyr421, Tyr466, and Tyr482, leading to cortactin interaction with F-actin and Arp2/3. Erk can also phosphorylate cortactin on Ser405 and Ser418, causing recruitment of additional actin nucleation-promoting factors (NPFs), most importantly N-WASP, to the Arp2/3/ complex. Phosphorylated cortactin thus recruits and stabilizes Arp2/3 complex on preexisting F-actin, providing a nucleation site for a new actin branch to form. Cortactin overexpression in tumor cells enhances invasiveness as a result of highly active lamellipodia formation, also known as invadopodia. Cortactin contains an N-terminal aspartic acid-aspartic acid-tryptophan (DDW) sequence (a.a. 29-31 in chicken; a.a. 20-22 in human & mouse) that mediates its interaction with Arp2/3, followed by six and a half repeat sequences (a.a. 80-125, 126-162, 163-199, 200-236, 237-273, 274-310, 311-333 in chicken; a.a. 80-116, 117-153, 154-190, 191-227, 228-264, 265-301, 302-324 in human and mouse) that mediates F-actin interaction, a proline-rich coiled coil domain (a.a. 349-410 in chicken; a.a. 348-401 in human and mouse) that mediates Src SH3 domain interaction, and a C-terminal SH3 domain (a.a. 505-563 in chicken; a.a. 492-550 in human a.a. 488-546 in mouse) that contains Erk phosphorylation sites and is important for cortactin interaction with NPFs other than Arp2/3.

Immunogen

GST-tagged recombinant human cortactin fragment comprising cortactin repeats 3-6.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Purified mouse IgG2b in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl with 0.05% sodium azide.

Protein G purified.

Format: Purified

Quality

Evaluated by Western Blotting in 3T3/A31 cell lysate.

Western Blotting Analysis: A 1:1,000 dilution of this antibody detected cortactin in 10 µg of 3T3/A31 cell lysate.

Specificity

Clone 5G3.2 epitope lies within the region containing cortactin repeats 3-6. Reactivity toward human spliced isoforms 2&3 that lack cortactin repeat 6 is possible, but has not been confirmed.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~75 kDa observed. Target band size appears larger than the calculated molecular weights of 61.59/70.96/57.47 kDa (human isoform 1/2/3) and 61.25 kDa (mouse) due to posttranslational modification. Uncharacterized bands may be observed in some lysate(s).