MA5-14831

antibody from Invitrogen Antibodies

Targeting: STK11 LKB1, PJS

Antibody data

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Validation data

STK11 protein structure - MA5-14831 shown in red.

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.

Supportive validation

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.

Supportive validation

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).