Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [3]
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Validation data
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- Product number
- MA5-14831 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- LKB1 Monoclonal Antibody (E.425.6)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- E.425.6
- Vial size
- 100 µL
- Concentration
- 24 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of LKB1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR870103_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of LKB1 was performed by loading 30 µg of A-431 Wild Type (Lane 1), A-431 CAS9 (Lane 2) and A-431 LKB1 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-LKB1 Monoclonal Antibody (E.425.6) (Product # MA5-14831, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to LKB1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of LKB1 was achieved by transfecting A-431 with LKB1 specific siRNAs (Silencer® select Products # s532020, s501375). Western blot analysis (Fig. a) was performed using whole cell extracts from the LKB1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with LKB1 Monoclonal Antibody (E.425.6) (Product # MA5-14831, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to LKB1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-LKB1 Monoclonal Antibody (E.425.6) (Product # MA5-14831) and a 55 kDa band corresponding to LKB1 was observed in all the tested cell models. Modified whole cell lysate (1% SDS) (30 µg lysate) of COS-7 (Lane 1), Caki-1 (Lane 2), K-562 (Lane 3), A-431 (Lane 4) and U-2 OS (Lane 5) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blots were probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).