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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
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- Product number
- 702178 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ITGA5 Recombinant Rabbit Monoclonal Antibody (12H40L20)
- Antibody type
- Monoclonal
- Antigen
- Other
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 12H40L20
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on Membrane enriched extracts (30 µg lysate) of U-87 MG (Lane 1), U-937 (Lane 2), THP-1 (Lane 3) and HeLa (Lane 4). The blots were probed with Anti-Integrin alpha 5 beta 1 Recombinant Rabbit Monoclonal Antibody (Product # 702178, 2.5 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:4000 dilution). A ~120 kDa band corresponding to Integrin alpha 5 beta 1 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 by wet transfer method. 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).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of ITGA5 was achieved by transfecting HeLa cells with ITGA5 specific validated siRNA (Silencer® select Cat # s7547). Western blot analysis was performed using membrane extracts from the ITGA5 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-ITGA5 Monoclonal Antibody (Product # 702178, 1-3 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to ITGA5.
