Validated using a knockout cell line
Recombinant
RabMAb

Anti-Integrin linked ILK antibody [EP1593Y] (ab52480)

Overview

  • Product name
    Anti-Integrin linked ILK antibody [EP1593Y]
    See all Integrin linked ILK primary antibodies
  • Description
    Rabbit monoclonal [EP1593Y] to Integrin linked ILK
  • Host species
    Rabbit
  • Specificity
    This antibody reacts with Integrin linked ILK. The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
  • Tested applications
    Suitable for: WB, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt or ICC
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human, African green monkey
  • Immunogen

    Synthetic peptide within Human Integrin linked ILK aa 400 to the C-terminus. The exact sequence is proprietary.
    Database link: Q13418

  • Positive control
    • Human heart muscle tissue and COS cell lysate
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EP1593Y
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab52480 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
WB 1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).

For unpurified use at 1/2000 - 1/10000 dilution. 

IP 1/40.

For unpurified use at 1/30 dilution. 

IHC-P 1/100.

See IHC antigen retrieval protocols.
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

  • Application notes
    Is unsuitable for Flow Cyt or ICC.
  • Target

    • Function
      Receptor-proximal protein kinase regulating integrin-mediated signal transduction. May act as a mediator of inside-out integrin signaling. Focal adhesion protein part of the complex ILK-PINCH. This complex is considered to be one of the convergence points of integrin- and growth factor-signaling pathway. Could be implicated in mediating cell architecture, adhesion to integrin substrates and anchorage-dependent growth in epithelial cells. Phosphorylates beta-1 and beta-3 integrin subunit on serine and threonine residues, but also AKT1 and GSK3B.
    • Tissue specificity
      Highly expressed in heart followed by skeletal muscle, pancreas and kidney. Weakly expressed in placenta, lung and liver.
    • Sequence similarities
      Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.
      Contains 5 ANK repeats.
      Contains 1 protein kinase domain.
    • Domain
      A PH-like domain is involved in phosphatidylinositol phosphate binding.
    • Post-translational
      modifications
      Autophosphorylated on serine residues.
    • Cellular localization
      Cell junction > focal adhesion. Cell membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • 59 kDa serine/threonine protein kinase antibody
      • 59 kDa serine/threonine-protein kinase antibody
      • DKFZp686F1765 antibody
      • Epididymis secretory protein Li 28 antibody
      • HEL S 28 antibody
      • ILK 1 antibody
      • ILK 2 antibody
      • ILK antibody
      • ILK-1 antibody
      • ILK-2 antibody
      • ILK_HUMAN antibody
      • ILK1 antibody
      • ILK2 antibody
      • Integrin linked kinase 2 antibody
      • Integrin linked Kinase antibody
      • Integrin linked protein kinase antibody
      • Integrin-linked protein kinase antibody
      • p59 antibody
      • p59ILK antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: Integrin linked ILK knockout HAP1 cell lysate (20 µg)
      Lane 3: HEK293 cell lysate (20 µg)
      Lane 4: K562 cell lysate (20 µg) 
      Lanes 1 - 4: Merged signal (red and green). Green - ab52480 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.

      Unpurified ab52480 was shown to specifically react with Integrin linked ILK when Integrin linked ILK knockout samples were used. Wild-type and Integrin linked ILK knockout samples were subjected to SDS-PAGE. Unpurified ab52480 and ab8245 (loading control to GAPDH) were both diluted 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling Integrin linked ILK with Purified ab52480 at 1:100 dilution (7.13 μg/ml). Heat mediated antigen retrieval was performed using citrate (pH 6.0)

    • All lanes : Anti-Integrin linked ILK antibody [EP1593Y] (ab52480) at 1/1000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
      Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates with 5% NFDM/TBST
      Lane 3 : Mouse heart lysates with 5% NFDM/TBST
      Lane 4 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates with 5% NFDM/TBST
      Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates with 5% NFDM/TBST

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 51 kDa
      Observed band size: 51 kDa

    • ab52480 (purified) at 1:40 dilution (2µg) immunoprecipitating Integrin linked ILK in HeLa whole cell lysate.
      Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
      Lane 2 (+): ab52480 & HeLa whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52480 in HeLa whole cell lysate
      For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    • Overlay histogram showing HEK293 cells stained with unpurified ab52480 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (unpurified ab52480, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor® 488 (IgG; H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    • Anti-Integrin linked ILK antibody [EP1593Y] (ab52480) at 1/1000 dilution (Purified) + COS-1 (African green monkey kidney fibroblast-like) whole cell lysates at 15 µg with 5% NFDM/TBST

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 51 kDa
      Observed band size: 51 kDa

    • Anti-Integrin linked ILK antibody [EP1593Y] (ab52480) at 1/2000 dilution (unpurified) + COS-1 cell lysate at 10 µg

      Secondary
      Goat anti-rabbit HRP labeled at 1/2000 dilution

      Predicted band size: 51 kDa
      Observed band size: 51 kDa

    • Anti-Integrin linked ILK antibody [EP1593Y] (ab52480) (unpurified) + PC12

      Predicted band size: 51 kDa

    • Immunohistochemical analysis of paraffin-embedded human heart muscle using unpurified ab52480 at a 1/50 dilution.

    • All lanes : Anti-Integrin linked ILK antibody [EP1593Y] (ab52480) at 1/2000 dilution (unpurified)

      Lane 1 : Rat Brain
      Lane 2 : Rat Heart
      Lane 3 : Rat Kidney
      Lane 4 : Rat Spleen

      Predicted band size: 51 kDa


      Exposure time: 3 minutes

    References

    This product has been referenced in:
    • Bai L  et al. Effect of Huayu Tongluo Herbs on Reduction of Proteinuria via Inhibition of Wnt/ß-Catenin Signaling Pathway in Diabetic Rats. Evid Based Complement Alternat Med 2017:3054145 (2017). WB ; Rat . Read more (PubMed: 28656052) »
    • Lu W  et al. Expression of integrin-linked kinase improves cardiac function in a swine model of myocardial infarction. Exp Ther Med 13:1868-1874 (2017). Read more (PubMed: 28565779) »

    See all 8 Publications for this product

    Customer reviews and Q&As

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    Unfortunately, we don't know the concentration ourselves: This is a tissue culture supernatant, and it is impossible to measure the concentration due to all the proteins in this solution.

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