09-142

Anti-phospho-ACK1 (Tyr284) Antibody

Upstate^®, from rabbit

• Synonym(s): TNK2, tyrosine kinase non-receptor 2, p21cdc42Hs, activated Cdc42-associated kinase 1, activated p21cdc42Hs kinase
• UNSPSC Code: 12352203
• eCl@ss: 32160702
• NACRES: NA.41

Properties

• biological source: rabbit
• Quality Level: 100
• antibody form: affinity purified immunoglobulin
• clone: polyclonal
• purified by: affinity chromatography
• species reactivity: human
• manufacturer/tradename: Upstate^®
• technique(s): dot blot: suitable; inhibition assay: suitable; western blot: suitable
• isotype: IgG
• NCBI accession no.: NM_001010938.1; NM_005781.4
• UniProt accession no.: Q07912
• shipped in: wet ice
• target post-translational modification: phosphorylation (pTyr284)
• Gene Information: human ... TNK2(10188)

Description

General description

Activated CDC42 kinase 1 (UniProt: Q07912; also known as ACK-1, Tyrosine kinase non-receptor protein 2) is encoded by the TNK2 (also known as ACK1) gene (Gene ID: 10188) in human. ACK-1 is a non-receptor tyrosine-protein kinase that is implicated in cell spreading and migration, cell survival, cell growth, and proliferation. It transduces extracellular signals to cytosolic and nuclear effectors. It is known to phosphorylate AKT1 (on tyrosine 176), androgen receptor (AR; on tyrosine 267), and WWOX (on tyrosine 287). It is also reported to phosphorylate MCF2, thereby enhancing its activity as a guanine nucleotide exchange factor (GEF) toward Rho family proteins. It has also been implicated in trafficking and clathrin-mediated endocytosis through binding to epidermal growth factor receptor (EGFR) and clathrin. It binds to both poly- and mono-ubiquitin and regulates ligand-induced degradation of EGFR, thereby contributing to the accumulation of EGFR at the limiting membrane of early endosomes. Its protein kinase domain is localized in amino acids 126-385 and its SH3 domain resides in amino acids 388-448. It also contains a SAM-like domain that is reported to be essential for NEDD4-mediated ubiquitination and promotes its membrane localization and dimerization to allow for autophosphorylation . It undergoes phosphorylation on tyrosine 284 and the phosphorylated form shows a significant increase in expression in prostate cancer breast cancers during progressive stages. (Ref.: Mahajan, K., et al. (2010). PLoS One. 5(3); e9646; Prieto-Echagüe, V., et al. (2010). BMC Biochem. 22; 42; van der Horst, E., et al. (2005). Proc. Natl. Acad. Sci. USA. 102(44); 15901-15906).

Specificity

This rabbit polyclonal antibody detects Activated CDC42 kinase 1 (ACK1) phosphorylated on tyrosine 284.

Immunogen

KLH-conjugated linear peptide corresponding to 9 amino acids surrounding human phospho ACK1 (Y284).

Application

Anti-phospho-ACK1 (Tyr284), Cat. No. 09-142, is a rabbit polyclonal antibody that detects ACK1 phosphorylated on tyrosine 284 and is tested for use in Dot Blot, Peptide Inhibition Assay, and Western Blotting.




Quality Control Testing

Evaluated by Western Blotting in A431 cell lysate.Western Blotting Analysis: A 1:250 dilution of this antibody detected phospho-ACK1(Tyr284) in A431 cell lysate.


Tested ApplicationsDot Blot Analysis: Anti-phospho-ACK1 (Tyr284)) antibody 09-142 detected Phospho-ACK1 (Tyr284) peptide, but not the unmodified ACK1 peptide.Peptide Inhibition Assay: Target band detection in a lysate from A431 cells was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Quality

Routinely evaluated by immunoblot.

Target description

~160 kDa observed; 114.57 kDa calculated. Uncharacterized bands may be observed in some lysate(s).

Physical form

Purified rabbit polyclonal antibody in buffer containing 0.014 M phosphate buffer, pH 7.6, 0.175 M NaCl with 0.07% sodium azide with 30% glycerol.

Storage and Stability

Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Safety Information

• Storage Class Code: 10 - Combustible liquids
• WGK: WGK 2