Explore
Validate
Learn
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- 711291 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ILK Recombinant Polyclonal Antibody (14HCLC)
- Antibody type
- Polyclonal
- Antigen
- Other
- Description
- This antibody is predicted to react with Monkey, Rat, Rabbit and Horse
- Antibody clone number
- 14HCLC
- Concentration
- 0.5 mg/mL
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), U-87 MG (Lane 2), K-562 (Lane 3) and HEK-293 (Lane 4). The blots were probed with Anti-Integrin-linked kinase Recombinant Rabbit Polyclonal Antibody (Product # 711291, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). 51 kDa and 36 kDa bands corresponding to Integrin-linked kinase was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- For immunofluorescence analysis, A549 cells were fixed and permeabilized for detection of endogenous Integrin-linked kinase (ILK) using Anti- Integrin-linked kinase (ILK) Recombinant Rabbit Polyclonal Antibody (Product # 711291, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Integrin-linked kinase (ILK) protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating localization of Integrin-linked kinase (ILK) in the cell membrane and focal adhesion junctions. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
