Anti-ACK1 antibody (ab65108)
Key features and details
- Rabbit polyclonal to ACK1
- Suitable for: ICC/IF, WB
- Reacts with: Rat, Human
- Isotype: IgG
Overview
-
Product name
Anti-ACK1 antibody
See all ACK1 primary antibodies -
Description
Rabbit polyclonal to ACK1 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Mouse, Cow, Dog, Rhesus monkey
-
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 1000 to the C-terminus of Human ACK1.
Read Abcam's proprietary immunogen policy (Peptide available as ab74487.) -
Positive control
- This antibody gave a positive signal in the following Whole Cell Lysates: HeLa, HepG2, PC12, SHSY-5Y, T24/83, HT 1080
-
General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
-
Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab65108 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes ICC/IF Use a concentration of 1 µg/ml. WB Use a concentration of 1 µg/ml. Detects a band of approximately 120 kDa (predicted molecular weight: 114 kDa). Target
-
Function
Non-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 specificity
The 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 similarities
Belongs 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. -
Domain
The 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. -
Post-translational
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 localization
Cell 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
-
Database links
- Entrez Gene: 280710 Cow
- Entrez Gene: 10188 Human
- Entrez Gene: 51789 Mouse
- Entrez Gene: 303882 Rat
- Omim: 606994 Human
- SwissProt: Q17R13 Cow
- SwissProt: Q07912 Human
- SwissProt: O54967 Mouse
see all -
Alternative names
- Acetate kinase 1 antibody
- Acetokinase 1 antibody
- ACK 1 antibody
see all
Images
-
Western blot - Anti-ACK1 antibody (ab65108)All lanes : Anti-ACK1 antibody (ab65108) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 4 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 5 : T24/83 (Human bladder carcinoma) Whole Cell Lysate
Lane 6 : HT 1080 (Human fibrosarcoma) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 114 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutes -
Immunocytochemistry/ Immunofluorescence - Anti-ACK1 antibody (ab65108)ICC/IF image of ab65108 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab65108, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Protocols
Datasheets and documents
References (2)
ab65108 has been referenced in 2 publications.
- 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). PubMed: 26677978
- Wilson C et al. Overcoming EMT-associated resistance to anti-cancer drugs via Src/FAK pathway inhibition. Oncotarget 5:7328-41 (2014). WB ; Human . PubMed: 25193862
Images
-
Western blot - Anti-ACK1 antibody (ab65108)All lanes : Anti-ACK1 antibody (ab65108) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
Lane 4 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate
Lane 5 : T24/83 (Human bladder carcinoma) Whole Cell Lysate
Lane 6 : HT 1080 (Human fibrosarcoma) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 114 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?
Exposure time: 4 minutes
-
Immunocytochemistry/ Immunofluorescence - Anti-ACK1 antibody (ab65108)ICC/IF image of ab65108 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab65108, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
