This is a knockout-validated antibody summary, based on the publication "Integrin-Linked Kinase (ILK) Deletion Disrupts Oligodendrocyte Development by Altering Cell Cycle", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: ILK1

Catalog number: 3856

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to the carboxy terminus of human ILK1. Reacts with human, mouse, rat, hamster, monkey, zebrafish, bovine, and pig. Suitable for western blot and immunoprecipitation. The antibody may cross-react with ILK2.

Western blot | Immunohistochemistry

Brains and spinal cords from control floxed and ILK cKO mice. WB: Neural tissue was homogenized in RIPA buffer (25 mm Tris-HCl, pH 7.5, 150 mm NaCl, 1 mm EDTA, 1% NP-40, 0.1% SDS, 0.1% DOC, R0278; Sigma-Aldrich) supplemented with protease inhibitor (complete mini, #04693124001; Roche) and phosphatase inhibitor (#524625; Calbiochem). IHC: Transgenic pups were anesthetized and perfused transcardially with 4% paraformaldehyde (PFA). Brains and spinal cords were dissected out and postfixed in the same fixative overnight, followed by cryoprotection with 30% sucrose and blocked in OCT (Sakura Finetek).

5% normal donkey serum (NDS) for 1 h.

WB: 1:1000

IHC: for 2 h at room temperature or overnight at 4°C.

WB: 1:10,000 dilution near-infrared fluorescent secondary antibodies (LI-COR).

IHC: Secondary antibodies (Jackson ImmunoResearch Laboratories) were either fluorescently conjugated or biotinylated (for DAB reaction or streptavidin reaction) and diluted in 5% NDS-PBS, 0.3% Triton X-100. The incubation time ranged from 60 to 90 min at room temperature.

WB: Proteins were visualized on an Odyssey Infrared Imaging System (LI-COR).

Immunohistochemistry of ILK shows that ILK was detected in CC1+ oligodendrocytes (arrows), to a lesser extent in NG2+ OPCs (arrowheads) in control fl/fl mice. Deletion was evident in CC1+ and NG2+ oligodendrocytes in ILK cKO mice. Western blot at P0 shows reduced ILK in brains and spinal cords from ILK cKO mice. Please see Figure 1c,d in the article [1].