Product nameAnti-ACK1 (phospho Y284) antibody
See all ACK1 primary antibodies
DescriptionRabbit polyclonal to ACK1 (phospho Y284)
Specificityab74091 detects ACK1 only when phosphorylated at tyrosine 284.
Tested applicationsSuitable for: WB, ELISA, IHC-P, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse
Synthetic phosphopeptide derived from human ACK1 around the phosphorylation site of tyrosine 284 (D-H-YP-V-M).
- Human breast carcinoma tissue, A549 cells, and HepG2 cell extracts treated with EGF (200ng/ml, 30mins).
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 50% Glycerol, 0.87% Sodium chloride, PBS
Without Mg2+ and Ca2+
Concentration information loading...
PurityImmunogen affinity purified
Purification notesab74091 was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific phosphopeptide. The antibody against non-phosphopeptide was removed by chromatography using non-phosphopeptide corresponding to the phosphorylation site.
Our Abpromise guarantee covers the use of ab74091 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/500 - 1/1000. Detects a band of approximately >117 kDa (predicted molecular weight: 114 kDa).|
|IHC-P||1/50 - 1/100.|
|ICC/IF||1/500 - 1/1000.|
FunctionNon-receptor tyrosine kinase that regulates the activity of a number of proteins by tyrosine phosphorylation especially proteins critical for cell survival, cell growth, and proliferation. Activates AKT1 by phosphorylating it on 'Tyr-176' resulting in its activation. Phosphorylates AR on 'Tyr-267' and 'Tyr-363' and promotes its recruitment to the androgen-responsive enhancers (AREs). Phosphorylates WWOX on 'Tyr-287'. Downstream effector of CDC42 which mediates CDC42-dependent cell migration via phosphorylation of BCAR1. Binds to both poly- and mono-ubiquitin and regulates ligand-induced degradation of EGFR. Participates in clathrin-mediated endocytosis. May be involved both in adult synaptic function and plasticity and in brain development.
Tissue specificityThe Tyr-284 phosphorylated form shows a significant increase in expression in breast cancers during the progressive stages i.e. normal to hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC) and lymph node metastatic (LNMM) stages. It also shows a significant increase in expression in prostate cancers during the progressive stages.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family.
Contains 1 CRIB domain.
Contains 1 protein kinase domain.
Contains 1 SH3 domain.
Contains 1 UBA domain.
DomainThe EBD (EGFR-binding domain) domain is necessary for interaction with EGFR.
The SAM-like domain is necessary for NEDD4-mediated ubiquitination. Promotes membrane localization and dimerization to allow for autophosphorylation.
The UBA domain binds both poly- and mono-ubiquitin.
modificationsAutophosphorylation regulates kinase activity. Phosphorylation on Tyr-518 is required for interaction with SRC.
Ubiquitinated by NEDD4. Its EGF-induced degradation is EGFR activation-dependent and is processed by lysosomes, not proteasomes.
Cellular localizationCell membrane. Nucleus. Endosome. Cell junction > adherens junction. Cytoplasmic vesicle membrane. The Tyr-284 phosphorylated form is expressed both in the membrane and nucleus. Co-localizes with EGFR on the endosomes.
- Information by UniProt
- Acetate kinase 1 antibody
- Acetokinase 1 antibody
- ACK 1 antibody
All lanes : Anti-ACK1 (phospho Y284) antibody (ab74091) at 1/500 dilution
Lane 1 : HepG2 cell extracts
treated with EGF (200ng/ml, 30mins)
Lane 2 : HepG2 cell extracts
treated with EGF (200ng/ml, 30mins) with immunising peptide at 10 µg
Lysates/proteins at 30 µg per lane.
Predicted band size: 114 kDa
Observed band size: >117 kDa why is the actual band size different from the predicted?
ab74091 at 1/500 dilution staining ACK1 in A549 cells by Immunofluorescence, in the absence or presence of the immunising peptide.
ab74091 at 1/50 dilution staining ACK1 in human breast carcinoma by Immunohistochemistry, Paraffin-embedded tissue, in the absence or presence of the immunising peptide.
This product has been referenced in:
- Shen R et al. The clinicopathological significance of ubiquitin-conjugating enzyme E2C, leucine-rich repeated-containing G protein-coupled receptor, WW domain-containing oxidoreductase, and vasculogenic mimicry in invasive breast carcinoma. Medicine (Baltimore) 98:e15232 (2019). Read more (PubMed: 31008954) »
- Maxson JE et al. Identification and Characterization of Tyrosine Kinase Nonreceptor 2 Mutations in Leukemia through Integration of Kinase Inhibitor Screening and Genomic Analysis. Cancer Res 76:127-38 (2016). Read more (PubMed: 26677978) »