Overview

  • Product nameAnti-ACK1 antibody
    See all ACK1 primary antibodies
  • Description
    Rabbit polyclonal to ACK1
  • Tested applicationsSuitable 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.

    (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

Properties

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

  • 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.
  • Post-translational
    modifications
    Autophosphorylation 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
  • Database links
  • Alternative names
    • Acetate kinase 1 antibody
    • Acetokinase 1 antibody
    • ACK 1 antibody
    • ACK antibody
    • ACK-1 antibody
    • ACK1 antibody
    • ACK1_HUMAN antibody
    • Activated Cdc42 associated kinase 1 antibody
    • Activated CDC42 kinase 1 antibody
    • Activated p21cdc42Hs kinase antibody
    • FLJ44758 antibody
    • FLJ45547 antibody
    • p21cdc42Hs antibody
    • TNK 2 antibody
    • TNK2 antibody
    • Tyrosine kinase non receptor 2 antibody
    • Tyrosine kinase non receptor protein 2 antibody
    • Tyrosine kinase non-receptor protein 2 antibody
    see all

Anti-ACK1 antibody images

  • 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
    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
  • 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.

References for Anti-ACK1 antibody (ab65108)

This product has been referenced in:
  • Wilson C  et al. Overcoming EMT-associated resistance to anti-cancer drugs via Src/FAK pathway inhibition. Oncotarget 5:7328-41 (2014). WB ; Human . Read more (PubMed: 25193862) »

See 1 Publication for this product

Product Wall

Thank you for contacting us.
I have double checked the western blots obtained for this particular lot, and the results looked good.
The lysis buffer we used to prepare our cell lysates is the following:
300 ul RIPA buffer
100 ul 10x...

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Thank you for taking time to submit your protocol and for contacting us. I am sorry to hear this antibody is not providing satisfactory results.
The details provided will enable us to investigate this case and will provide us with vital informatio...

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% TritonX100 in PBS
Username

Dr. Kirk McManus

Verified customer

Submitted Nov 20 2009

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"