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Integrin linked ILK Recombinant Rabbit Monoclonal Antibody [SC68-04]

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Specification

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Catalog# ET1610-76

Integrin linked ILK Recombinant Rabbit Monoclonal Antibody [SC68-04]

Application

  • WB

  • IF-Cell

  • IHC-P

  • IP

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Integrin linked ILK Recombinant Rabbit Monoclonal Antibody [SC68-04]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within human Integrin linked ILK aa 410-452.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, IP, FC

Molecular Weight

Predicted band size: 51 kDa

Positive Control

HeLa cell lysate, A549 cell lysate, NCI-H1299 cell lysate, NIH/3T3, SH-SY5Y, rat brain tissue, Jurkat.

Conjugation

unconjugated

Clone Number

SC68-04

RRID

AB_3069961

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

  • IP

  • 1-2μg/sample

Target

Function

Integrins are heterodimers composed of non-covalently associated transmembrane a and b subunits. The 16 a and 8 b subunits heterodimerize to produce more than 20 different receptors. Most integrin receptors bind to ligands that are components of the extracellular matrix. Certain integrins can also bind to soluble ligands such as Fibrinogen, or to counterreceptors on adjacent cells, such as the intracellular adhesion molecules (ICAMs), leading to aggregation of cells. In addition to mediating cell adhesion and cytoskeletal organization, integrins function as signaling receptors. Signals transduced by integrins play a role in many biological processes, including cell growth, differentiation, migration and apoptosis. ILK (integrin-linked kinase) was identified as a serine/threonine kinase that phosphorylates b1 and b3 integrins. ILK expression has been shown to be reduced in response to Fibronectin, a known integrin ligand. Overexpression of ILK was shown to upregulate the Fibronectin matrix assembly in epithelial cells, indicating a potential role for ILK in cell growth, cell survival and tumorigenesis.

Background References

1. Hoshino A et al. Tumour exosome integrins determine organotropic metastasis. Nature 527:329-35 (2015).

2. Rottmar M et al. Interference with the contractile machinery of the fibroblastic chondrocyte cytoskeleton induces re-expression of the cartilage phenotype through involvement of PI3K, PKC and MAPKs. Exp Cell Res 320:175-87 (2014).

Sequence Similarity

Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family.

Tissue Specificity

Highly expressed in heart followed by skeletal muscle, pancreas and kidney. Weakly expressed in placenta, lung and liver.

Post-translational Modification

Autophosphorylated on serine residues.

Subcellular Location

Cell junction, Cell membrane, Cytoplasm.

UNIPROT

SwissProt: Q13418 Human

SwissProt: O55222 Mouse

SwissProt: Q99J82 Rat

Synonyms

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

Expand

Images

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of Integrin linked ILK on different lysates with Rabbit anti-Integrin linked ILK antibody (<a href="/products/ET1610-76" style="font-weight: bold;text-decoration: underline;">ET1610-76</a>) at 1/1,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-Integrin linked ILK KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 51 kDa<br />Observed band size: 51 kDa<br /><br />Exposure time: 1 minute; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-76" style="font-weight: bold;text-decoration: underline;">ET1610-76</a>) at 1/1,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of Integrin linked ILK on different lysates with Rabbit anti-Integrin linked ILK antibody (ET1610-76) at 1/1,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-Integrin linked ILK KD cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 1 minute; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-76) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Integrin linked ILK on different lysates with Rabbit anti-Integrin linked ILK antibody (<a href="/products/ET1610-76" style="font-weight: bold;text-decoration: underline;">ET1610-76</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: A549 cell lysate<br />Lane 3: NCI-H1299 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 51 kDa<br />Observed band size: 51 kDa<br /><br />Exposure time: 25 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1610-76" style="font-weight: bold;text-decoration: underline;">ET1610-76</a>) at 1/1,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Integrin linked ILK on different lysates with Rabbit anti-Integrin linked ILK antibody (ET1610-76) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: A549 cell lysate
    Lane 3: NCI-H1299 cell lysate

    Lysates/proteins at 20 µg/Lane.

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

    Exposure time: 25 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-76) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • ICC staining of Integrin linked ILK in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1610-76" style="font-weight: bold;text-decoration: underline;">ET1610-76</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Integrin linked ILK in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-76, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • ICC staining of Integrin linked ILK in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (<a href="/products/ET1610-76" style="font-weight: bold;text-decoration: underline;">ET1610-76</a>, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor&reg;488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

    ICC staining of Integrin linked ILK in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-76, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

  • Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Integrin linked ILK antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1610-76" style="font-weight: bold;text-decoration: underline;">ET1610-76</a>, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Integrin linked ILK antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-76, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of Integrin linked ILK was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1610-76" style="font-weight: bold;text-decoration: underline;">ET1610-76</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Integrin linked ILK was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1610-76, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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Citation

Products with the same target and pathway

metafield image

Integrin linked ILK Rabbit Polyclonal Antibody

Application: WB,IF-Cell,IHC-P

Reactivity: Human,Mouse,Rat

Conjugate: unconjugated