Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
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- Product number
- MA5-15282 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- FAK Monoclonal Antibody (4A9D6)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15282 targets FAK in indirect ELISA, WB applications and shows reactivity with Human samples. The MA5-15282 immunogen is purified recombinant fragment of FAK expressed in E. Coli.
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 4A9D6
- Vial size
- 100 µL
- Concentration
- Conc. Not Determined
- 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 of FAK using a FAK monoclonal antibody (Product # MA5-15282) against a truncated FAK recombinant protein.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of FAK using a FAK monoclonal antibody (Product # MA5-15282) against a truncated FAK recombinant protein.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- CRISPR-Cas9 mediated genome editing ofFAK (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, AssayID CRISPR627416_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of FAK was performed by loading 30 µg of A549 Cas9 (Lane 1) and A549 Cas9 cells transduced with FAK Lentiviral sgRNA (Lane 2) membrane enriched extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with FAK Monoclonal Antibody (4A9D6) (Product # MA5-15282, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10,000 dilution). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076) using the iBright™ FL 1500 (Product # A44115). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toFAK (Fig (b)). An uncharacterized band was observed at ~35 kDa in both the samples.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-FAK Monoclonal Antibody (4A9D6) (Product # MA5-15282) and a120kDa band corresponding to Focal adhesion kinase 1 was observed across cell lines tested except U-937 which is reported to be negative for FAK expression. Additional uncharacterized bands (*) were observed across the cell lines tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), HCT 116 (Lane 2), A549 (Lane 3), A-431 (Lane 4), SH-SY5Y (Lane 5), K-562 (Lane 6) and U-937 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)