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Key Documents

09-142

Sigma-Aldrich

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

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About This Item

UNSPSC Code:
12352203
eCl@ss:
32160702
NACRES:
NA.41

biological source

rabbit

Quality Level

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.

UniProt accession no.

shipped in

wet ice

target post-translational modification

phosphorylation (pTyr284)

Gene Information

human ... TNK2(10188)

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

Storage Class Code

10 - Combustible liquids

WGK

WGK 2


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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A study of somatolactin actions by ectopic expression in transgenic zebrafish larvae.
Guohui Wan,King Ming Chan
Journal of Molecular Endocrinology null
Y Liu et al.
Oncogene, 29(22), 3208-3216 (2010-04-13)
Activation of androgen receptor (AR) may have a role in the development of castration-resistant prostate cancer. Two intracellular tyrosine kinases, Ack1 (activated cdc42-associated kinase) and Src, phosphorylate and enhance AR activity and promote prostate xenograft tumor growth in castrated animals.
Wnt4 enhances murine hematopoietic progenitor cell expansion through a planar cell polarity-like pathway.
Heinonen KM, Vanegas JR, Lew D, Krosl J, Perreault C
Testing null
Mathew C Halter et al.
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White biotechnology has made a positive impact on the chemical industry by providing safer, more efficient chemical manufacturing processes that have reduced the use of toxic chemicals, harsh reaction conditions, and expensive metal catalysts, which has improved alignment with the
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Biochemical and biophysical research communications, 314(2), 571-579 (2004-01-22)
Ras signals for the transformation of mammalian cells are apparently transduced through Rho GTPases. The Rho GTPase family member Cdc42 generates independent signals that regulate the rearrangement of the actin cytoskeleton and the transcription of genes. However, the molecular mechanism

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