Product Name:
ACK1-pY518
Product Number:
ab-pk512
Target Full Name: Activated CDC42 kinase 1
Target Alias: ACK; Activated Cdc42-associated kinase 1; Activated p21cdc42Hs kinase; Activated p21cdc42Hs kinase 1; Kinase ACK1; Non-receptor protein tyrosine kinase Ack; p21cdc42Hs; TNK2; Tyrosine kinase, non-receptor, 2; FLJ44758; FLJ45547; ENSG00000061938
Product Type Specific: ACK1 protein kinase phosphosite-specific antibody
Antibody Code: PK512
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y518
Protein UniProt: Q07912
Protein SigNET: ACK1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human ACK1 (TNK2) sequence peptide Cat. No.: PE-04AHK99
Antibody Immunogen Sequence: KPT(pY)DPV(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues K515 to V521; Just after the GTPase-binding domain. The major site of phosphorylation of ACK1.Y518 phosphorylation stimulates phosphotransferase activity. This is major site of phosphorylation of ACK1 by mass spectrometry studies (≥760 mass spectrometry reports recorded in PhosphoSitePlus). ACK1 is known to be phosphorylated at this site in vitro by ACK1 (TNK2).
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Weak immunoreactivity of a target-sized protein by Western blotting in HEK-293 cells. Medium immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Rhesus Macaque | Dog | Rat.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 120-130 kDa.
Antibody Specificity: High
Antibody Cross Reactivity: Weak cross-reactivity with a ~27 kDa protein in HEK-293 and HeLa cells.
Related Product 1: Ack1 (515-521) pY518 antibody blocking peptide (Cat. No.: PE-04AHK99)
Related Product 2: ACK1 Y284 phosphorylation antibody (Cat. No.: AB-PK511)
Related Product 3: ACK1 Y859+Y860 phosphorylation antibody (Cat. No.: AB-PK513)
Related Product 4: AckSubtide - Ack1 (TNK2) protein kinase substrate peptide
Scientific Background: ACK1 (TNK2) is a 140 kDa protein-tyrosine kinase of the TK group and Ack family. It is implicated in numerous roles, including: cell spreading and migration, cell survival, cell growth, and proliferation. It is a multi-domain cytoplasmic protein that includes an N-terminal Sterile alpha-motif (SAM) domain, a catalytic kinase domain, an SH3 domain, a Cdc42-binding (CRIB) domain, and a C-terminal ubiquitin-association (UBA) domain. It is indirectly activated by various ligands including growth factors (EGF, PDGF), and integrins, directly interacts with Cdc42. Phosphorylation at Tyr-284 in ACK1 increases its phosphotransferase activity. Its SH3 domain appears to play an autoinhibitory role. ACK1 appears to be a tumour suppressor protein (TSP), since cancer-related mutations in ACK1 in human tumours point to a loss of function of the protein kinase. The active form of the protein kinase normally acts to inhibit tumour cell proliferation. ACK1 is known to phosphorylate Akt1, AR, MCF2, WASL and WWOX. In this capacity ACK1 negatively regulates the tumour suppressor, WWOX, whilst activating the oncoproteins, Akt1 and AR. It can act as an epigenetic imprinter by regulating Histone H4 Tyr-88 phosphorylation. Mutations in this gene have been observed in ovarian, skin, and stomach tumours. This description may include information annotated by UniProt and/or Google AI.